Substituted diarylamines as MEK inhibitors

ABSTRACT

Diarylamines, such as 5-amide substituted diarylamines of formula (I) or formula (II) wherein A is hydroxy, C 1-6  alkoxy, or NR 6 OR 7 ; X is OR 12 , NR 13 R 12 , or NR 14 ; inhibitors of MEK and are useful in the treatment of a variety of proliferative disease states, such as conditions related to the hyperactivity of MEK, as well as diseases modulated by the MEK cascade.

This application is a 371 application of PCT/US01/07816 filed Mar. 12,2001, which claims the benefit of priority to U.S. provisionalapplication Ser. No. 60/189,714 filed Mar. 15, 2000 and U.S. provisionalapplication Ser. No. 60/210,205 filed Jun. 8, 2000.

The present invention relates to diarylamines, such as 5-amidesubstituted diarylamines, and methods of use thereof.

BACKGROUND OF THE INVENTION

Mitogen ERK Kinase (“MEK”) enzymes are dual specificity kinases involvedin, for example, immunomodulation, inflammation, and proliferativediseases such as cancer and restenosis.

Proliferative diseases are caused by a defect in the intracellularsignaling system, or the signal transduction mechanism of certainproteins. Defects include a change either in the intrinsic activity orin the cellular concentration of one or more signaling proteins in thesignaling cascade. The cell may produce a growth factor that binds toits own receptors, resulting in an autocrine loop, which continuallystimulates proliferation. Mutations or overexpression of intracellularsignaling proteins can lead to spurious mitogenic signals within thecell. Some of the most common mutations occur in genes encoding theprotein known as Ras, a G-protein that is activated when bound to GTP,and inactivated when bound to GDP. The above-mentioned growth factorreceptors, and many other mitogenic receptors, when activated, lead toRas being converted from the GDP-bound state to the GTP-bound state.This signal is an absolute prerequisite for proliferation in most celltypes. Defects in this signaling system, especially in the deactivationof the Ras-GTP complex, are common in cancers, and lead to the signalingcascade below Ras being chronically activated.

Activated Ras leads in turn to the activation of a cascade ofserine/threonine kinases. One of the groups of kinases known to requirean active Ras-GTP for its own activation is the Raf family. These inturn activate MEK (e.g., MEK₁ and MEK₂) which then activates the MAPkinase, ERK (ERK₁ and ERK₂). Activation of MAP kinase by mitogensappears to be essential for proliferation; constitutive activation ofthis kinase is sufficient to induce cellular transformation. Blockade ofdownstream Ras signaling, for example by use of a dominant negativeRaf-1 protein, can completely inhibit mitogenesis, whether induced fromcell surface receptors or from oncogenic Ras mutants. Although Ras isnot itself a protein kinase, it participates in the activation of Rafand other kinases, most likely through a phosphorylation mechanism. Onceactivated, Raf and other kinases phosphorylate MEK on two closelyadjacent serine residues, S²¹⁸ and S²²² in the case of MEK-1, which arethe prerequisite for activation of MEK as a kinase. MEK in turnphosphorylates MAP kinase on both a tyrosine, Y¹⁸⁵, and a threonineresidue, T¹⁸³, separated by a single amino acid. This doublephosphorylation activates MAP kinase at least 100-fold. Activated MAPkinase can then catalyze the phosphorylation of a large number ofproteins, including several transcription factors and other kinases.Many of these MAP kinase phosphorylations are mitogenically activatingfor the target protein, such as a kinase, a transcription factor, oranother cellular protein. In addition to Raf-1 and MEKK, other kinasesactivate MEK, and MEK itself appears to be a signal integrating kinase.Current understanding is that MEK is highly specific for thephosphorylation of MAP kinase. In fact, no substrate for MEK other thanthe MAP kinase, ERK, has been demonstrated to date and MEK does notphosphorylate peptides based on the MAP kinase phosphorylation sequence,or even phosphorylate denatured MAP kinase. MEK also appears toassociate strongly with MAP kinase prior to phosphorylating it,suggesting that phosphorylation of MAP kinase by MEK may require a priorstrong interaction between the two proteins. Both this requirement andthe unusual specificity of MEK are suggestive that it may have enoughdifference in its mechanism of action to other protein kinases thatselective inhibitors of MEK, possibly operating through allostericmechanisms rather than through the usual blockade of the ATP bindingsite, may be found.

It has been found that the compounds of the present invention areinhibitors of MEK and are useful in the treatment of a variety ofproliferative disease states, such as conditions related to thehyperactivity of MEK, as well as diseases modulated by the MEK cascade.

SUMMARY

The present invention provides compounds of formula I and II:

wherein

-   -   R₁ is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkoxy, halo, C₁₋₂ haloalkyl,or        CN;    -   R₃ and R₄ are each independently hydrogen, halo, C₁₋₂ haloalkyl,        C₁₋₈ alkyl, C₁₋₈ alkoxy, nitro, CN, or (O or        NH)_(k)—(CH₂)_(j)—R₉, where R₉ is hydrogen, hydroxy, CO₂H or        NR₁₀R₁₁;    -   j is 0 to 4;    -   k is 0 or 1;    -   R₁₀ and R₁₁ are each independently hydrogen or C₁₋₈ alkyl, or        together with the nitrogen to which they are attached form a 3-        to 10-member cyclic ring optionally containing one, two, or        three additional heteroatoms selected from the group consisting        of O, S, NH, and N—C₁₋₈ alkyl;    -   A is hydroxy, C₁₋₆ alkoxy, or NR₆OR₇;    -   R₆ is hydrogen, C₁₋₈ alkyl, (CO)—C₁₋₈ alkyl, phenyl, naphthyl,        phenyl(C₁₋₈ alkyl), or C₃₋₁₀ cycloalkyl;    -   R₇ is hydrogen, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀        cycloalkyl or C₃₋₁₀ cycloalkyl optionally containing a        heteroatom selected from the group consisting of O, S, and NR₉;    -   X is OR₁₂, NR₁₃R₁₂, or NR₁₄;

R₁₂ and R₁₃ are each independently hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, C₄₋₆ cycloalkyl, [(CH₂)_(n)Y(CH₂)_(m)]_(q)CH₃, phenyl,naphthyl, (C₁₋₆ alkyl)phenyl, —[(CH₂)_(n)Y(CH₂)_(m)]_(q)phenyl, C₂₋₆heteroaryl, (C₁₋₆ alkyl)C₂₋₆ heterocyclic radical, or[(CH₂)_(n)Y(CH₂)_(m)]_(q) C₂₋₆ heterocyclic radical;

Y is N or O;

R₁₄ taken with N is a 5- to 7-membered heterocyclic radical with between0 and 3 additional heteroatoms or heteroatom combinations in the ringselected from the group consisting of O, S, SO, SO₂, NH, and NMe; 0≦n,m≦6, n+m≦8, 1≦q≦5; and

wherein the above alkyl, alkenyl, alkynyl, heterocyclic radical, aryl,and cycloalkyl groups can be optionally substituted with between 1 and 4substituents independently selected from the group consisting ofhydroxy, C₁₋₄ alkyl, fluoro, chloro, iodo, bromo, amino, and C₁₋₄alkoxy, and NR_(a)R_(b);

wherein R_(a) and R_(b) are each independently selected from the groupconsisting of hydrogen and C₁₋₆ alkyl; and

the pharmaceutically acceptable salts thereof.

The invention also provides a pharmaceutical composition comprising acompound of formula I or II and a pharmaceutically acceptable carrier.

Additionally, the invention provides a method of treating aproliferative disease in a patient in need thereof comprisingadministering a therapeutically effective amount of a compound offormula I or II.

The invention also provides the use of a compound of formula I or II forthe manufacture of a medicament for the treatment of a proliferativedisease.

Furthermore, the invention provides methods of treating cancer,restenosis, psoriasis, autoimmune disease, atherosclerosis,osteoarthritis, rheumatoid arthritis, heart failure, chronic pain, andneuropathic pain in a patient in need thereof comprising administering atherapeutically effective amount of a compound of formula I or II.

The invention also provides the use of a compound of formula I or II forthe manufacture of a medicament for the treatment of cancer, restenosis,psoriasis, autoimmune disease, atherosclerosis, osteoarthritis,rheumatoid arthritis, heart failure, chronic pain, and neuropathic pain.

In addition, the invention provides a method for treating cancer in apatient in need thereof comprising administering a therapeuticallyeffective amount of a compound of formula I or II in combination withradiation therapy or at least one chemotherapeutic agent.

The invention also features synthetic intermediates and methodsdisclosed herein.

Other aspects of the invention are provided in the description,examples, and claims below.

DETAILED DESCRIPTION

The invention features diarylamine compounds, pharmaceuticalcompositions thereof, and methods of using such compounds andcompositions.

Certain terms are defined below and by their usage throughout thisdisclosure.

Alkyl groups, such as C₁₋₈ alkyl, include aliphatic chains (i.e.,hydrocarbyl or hydrocarbon radical structures containing hydrogen andcarbon atoms) with a free valence. Alkyl groups are understood toinclude straight chain and branched structures. Examples include methyl,ethyl, propyl, isopropyl, butyl, n-butyl, isobutyl, t-butyl, pentyl,isopentyl, 2,3-dimethylpropyl, hexyl, 2,3-dimethylhexyl,1,1-dimethylpentyl, heptyl, and octyl. The term “C₁₋₈ alkyl” includeswithin its definition the terms “C₁₋₆ alkyl” and “C₁₋₄ alkyl”.

Cycloalkyl groups, such as C₃₋₁₀ cycloalkyl, include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Theterm “C₃₋₁₀ cycloalkyl” includes within its definition the terms “C₄₋₆cycloalkyl”.

The term “halo” as used herein refers to fluoro, chloro, bromo, or iodo.

The term “haloalkyl” as used herein refers to a straight or branchedalkyl chain with 1, 2 or 3 halo atoms attached to it. The term “C₁₋₂haloalkyl” as used herein refers to a straight or branched alkyl chainhaving from one to two carbon atoms with 1, 2 or 3 halo atoms attachedto it. Typical C₁₋₂ haloalkyl groups include chloromethyl, 2-bromoethyl,difluoromethyl, trifluoromethyl and the like.

The term “alcoxy” as used herein refers to a straight or branched alkylchain attached to an oxygen atom. The term “C₁₋₈ alcoxy” as used hereinrefers to a straight or branched alkyl chain having from one to eightcarbon atoms attached to an oxygen atom. Typical C₁₋₈ alcoxy groupsinclude methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, pentoxyand the like. The term “C₁₋₈ alcoxy” includes within its definition theterms “C₁₋₆ alcoxy” and “C₁₋₄ alcoxy”.

Alkyl and cycloalkyl groups can be substituted with 1, 2, 3 or moresubstituents which are independently selected from hydroxy, alkyl, halo,amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy,arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy.Specific examples include fluoromethyl, hydroxyethyl,2,3-dihydroxyethyl, (2- or 3-furanyl)methyl, cyclopropylmethyl,benzyloxyethyl, (3-pyridinyl)methyl, (2- or 3-furanyl)methyl,(2-thienyl)ethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl,methoxymethyl, N-pyridinylethyl, diethylaminoethyl, andcyclobutylmethyl.

In some embodiments, each hydrocarbon radical above is optionallysubstituted with between 1 and 3 or more substituents independentlyselected from halo, hydroxyl or hydroxy, amino, (amino)sulfonyl, andNO₂. In another embodiment, each heterocyclic radical above isoptionally substituted with between 1 and 3 or more substituentsindependently selected from halo, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₃₋₄alkenyl, C₃₋₄ alkynyl, phenyl, hydroxyl or hydroxy, C₁–C₄ alkoxy amino,(amino)sulfonyl, and NO₂, wherein each substituent alkyl, cycloalkyl,alkenyl, alkynyl or phenyl is in turn optionally substituted withbetween 1 and 2 substituents independently selected from halo, C₁₋₂alkyl, hydroxyl or hydroxy, amino, and NO₂.

More general forms of substituted hydrocarbon radicals includehydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl,hydroxyaryl, and corresponding forms for the prefixes amino-, halo-,nitro-, alkyl-, phenyl-, cycloalkyl- and so on, or combinations ofsubstituents. According to formula I and II, therefore, substitutedalkyls include hydroxyalkyl, aminoalkyl, nitroalkyl, haloalkyl,alkylalkyl (branched alkyls, such as methylpentyl), (cycloalkyl)alkyl,phenylalkyl, alkoxyalkyl, alkylaminoalkyl, dialkylaminoalkyl, arylalkyl,aryloxyalkyl, arylalkyloxyalkyl, (heterocyclic radical)alkyl, and(heterocyclic radical)oxyalkyl.

R₁ thus includes hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl,hydroxycycloalkyl, hydroxyaryl, aminoalkyl, aminoalkenyl, aminoalkynyl,aminocycloalkyl, aminoaryl, alkylalkenyl, (alkylaryl)alkyl,(haloaryl)alkyl, (hydroxyaryl)alkynyl, and so forth. Similarly, R_(a)includes hydroxyalkyl and aminoaryl, and R_(b) includes hydroxyalkyl,aminoalkyl, and hydroxyalkyl(heterocyclic radical)alkyl.

Alkenyl groups are analogous to alkyl groups, but have at least onedouble bond (two adjacent sp² carbon atoms). Depending on the placementof a double bond and substituents, if any, the geometry of the doublebond may be entgegen (E), or zusammen (Z), cis, or trans. Similarly,alkynyl groups have at least one triple bond (two adjacent sp carbonatoms). Unsaturated alkenyl or alkynyl groups may have one or moredouble or triple bonds, respectively, or a mixture thereof; like alkylgroups, unsaturated groups may be straight chain or branched, and theymay be substituted as described both above for alkyl groups andthroughout the disclosure by example. Examples of alkenyls, alkynyls,and substituted forms include cis-2-butenyl, trans-2-butenyl, 3-butynyl,3-phenyl-2-propynyl, 3-(2′-fluorophenyl)-2-propynyl,3-methyl(5-phenyl)-4-pentynyl, 2-hydroxy-2-propynyl,2-methyl-2-propynyl, 2-propenyl, 4hydroxy-3-butynyl,3-(3-fluorophenyl)-2-propynyl, and 2-methyl-2-propenyl. In formulas Iand II, alkenyls and alkynyls can be C₂₋₄, C₂₋₆ or C₂₋₈, for example,and are preferably C₃₋₄ or C₃₋₈.

Heterocyclic radicals, which include but are not limited to heteroaryls,such as C₃₋₈ and C₂₋₆ heteroaryls, include: furyl, oxazolyl, isoxazolyl,thiophenyl, thiazolyl, pyrrolyl, imidazolyl, 1,3,4-triazolyl,tetrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indolyl, and theirnonaromatic counterparts. Further examples of heterocyclic radicalsinclude piperidyl, quinolyl, isothiazolyl, piperidinyl, morpholinyl,piperazinyl, tetrahydrofuryl, tetrahydropyrrolyl, pyrrolidinyl,octahydroindolyl, octahydrobenzothiofuranyl, and octahydrobenzofuranyl.Heterocyclic radicals may be substituted as described both above foralkyl groups and throughout the disclosure by example.

Heterocyclic radicals include heteroaryls such as substituted orUnsubstituted radicals of pyran, pyrazole, triazole, indazole, pyrazine,oxadiazole, oxathiadiazole; heterocycles also include heteroalkyls suchas substituted and unsubstituted radicals of tetrahydropyran,pyrrolidone, imidazoline, and tetrahydrothiophene.

The present invention includes pharmaceutically acceptable salts,amides, and esters of the disclosed compounds. The invention alsofeatures a pharmaceutically acceptable salt or C₁₋₈ ester of a disclosedcompound. For example, the disclosed alcohol compounds may form estershaving the structure obtained by replacing the H of a hydroxyl groupwith a —C(═O)C₁₋₇ acyl group.

The invention provides the disclosed compounds and closely related,pharmaceutically acceptable forms of the disclosed compounds, such assalts, esters, amides, hydrates or solvated forms thereof; masked orprotected forms; and racemic mixtures, or enantiomerically or opticallypure forms.

Pharmaceutically acceptable salts, esters, and amides includecarboxylate salts (e.g., C₁₋₈ alkyl, cycloalkyl, aryl, heteroaryl, ornon-aromatic heterocyclic), amino acid addition salts, esters, andamides which are within a reasonable benefit/risk ratio,pharmacologically effective, and suitable for contact with the tissuesof patients without undue toxicity, irritation, or allergic response.Representative salts include hydrobromide, hydrochloride, sulfate,bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate,stearate, laurate, borate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate,glucoheptonate, lactiobionate, and laurylsulfonate. These may includealkali metal and alkali earth cations such as sodium, potassium,calcium, and magnesium, as well as non-toxic ammonium, quaternaryammonium, and amine cations such as tetramethyl ammonium, methylamine,trimethylamine, and ethylamine. See, for example, S. M. Berge, et al.,“Pharmaceutical Salts,” J. Pharm. Sci., 1977, 66:1–19 which isincorporated herein by reference. Representative pharmaceuticallyacceptable amides of the invention include those derived from ammonia,primary C₁₋₆ alkyl amines and secondary di(C₁₋₆ alkyl) amines. Secondaryamines include 5- or 6-membered heterocyclic or heteroaromatic ringmoieties containing at least one nitrogen atom and optionally between 1and 2 additional heteroatoms. Preferred amides are derived from ammonia,C₁₋₃ alkyl primary amines, and di(C₁₋₂ alkyl)amines. Representativepharmaceutically acceptable esters of the invention include C₁₋₇ alkyl,C₅₋₇ cycloalkyl, phenyl, and phenyl(C₁₋₆)alkyl esters. Preferred estersinclude methyl esters.

The present invention includes compounds having one or more functionalgroups (e.g., hydroxyl, amino, or carboxyl) masked by a protectinggroup. Examples of protecting groups used to protect functional groupsand their preparation are disclosed by T. W. Green, “Protective Groupsin Organic Synthesis,” John Wiley & Sons, 1981. Choice of the protectinggroup used will depend upon the substituent to be protected and theconditions that will be employed in subsequent reaction steps whereinprotection is required, and is well within the knowledge of one ofordinary skill in the art. Protecting groups include, but are notlimited to, the list provided below.

Hydroxyl Protecting Groups

Hydroxyl protecting groups include: ethers, esters, and protection for1,2- and 1,3-diols. The ether protecting groups include: methyl,substituted methyl ethers, substituted ethyl ethers, substituted benzylethers, silyl ethers and conversion of silyl ethers to other functionalgroups.

Substituted methyl ethers include: methoxymethyl, methylthiomethyl,t-utylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyloxymethyl,p-ethoxybenzyloxymethyl, (4-methoxyphenoxy)methyl, guaiacolmethyl,t-butoxymethyl, 4-pentenyloxymethyl, siloxymethyl,2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl,bis(2-chloro-ethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl,tetrahydropyranyl, 3-bromotetrahydro-pyranyl, tetrahydrothiopyranyl,1-methoxycyclohexyl, 4-methoxytetrahydropyranyl,4-methoxytetrahydrothio-pyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxido, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl,1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, and2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-ethanobenzofuran-2-yl.

Substituted ethyl ethers include: 1-ethoxyethyl,1-(2,chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilyethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, andbenzyl.

Substituted benzyl ethers include: p-methoxybenzyl, 3,4-dimethoxybenzyl,o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl,p-cyanobenzyl, p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolylN-oxido, diphenylmethyl, p, p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenyl-methyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri-(p-methoxyphenyl)methyl,4-(4′-bromophenacyloxy)phenyldiphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-ylmethyl)bis(4′,4″-dimethoxyphenyl)-methyl,1,1-bis(4-methoxyphenyl)1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodithiolan-2-yl, and benzisothiazolyl S,S-dioxido.

Silyl ethers include: trimethylsilyl, triethylsilyl, triisopropylsilyl,dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl,t-butyldimethylsilyl, t-butyldiphenylsilyl, tribenzylsilyl,tri-pxylylsilyl, triphenylsilyl, diphenylmethylsilyl, andt-butylmethoxyphenylsilyl.

Ester Protecting Groups

Ester protecting groups include: esters, carbonates, assisted cleavage,miscellaneous esters, and sulfonates.

Examples of protective esters include: formate, benzoylformate, acetate,chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate,methoxyacetate, triphenylmethoxyacetate, phenoxyacetate,p-chlorophenoxyacetate, p-P-phenylacetate, 3-phenylpropionate,4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate, pivaloate,adamantoate,crotonate,4-methoxycrotonate, benzoate, p-phenylbenzoate,and 2,4,6-trimethylbenzoate(mesitoate).

Carbonates include: methyl, 9-fluorenylmethyl, ethyl,2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl,2-(triphenylphosphonio)ethyl, isobutyl, vinyl, allyl, p-nitrophenyl,benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl, and methyldithiocarbonate.

Examples of assisted cleavage protecting groups include: 2-iodobenzoate,4-azido-butyrate, 4-nitro4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzene-sulfonate, 2-(methylthiomethoxy)ethyl carbonate,4-(methylthiomethoxymethyl)benzoate, and2-(methylthiomethoxymethyl)benzoate.

In addition to the above classes, miscellaneous esters include:2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate (tigloate),o-(methoxycarbonyl)benzoate, p-P-benzoate, α-naphthoate, nitrate, alkylN,N,N′N′-tetramethylphosphorodiamidate, N-phenylcarbamate, borate,dimethylphosphinothioyl, and 2,4-dinitrophenylsulfenate.

Protective sulfates include: sulfate, methanesulfonate(mesylate),benzylsulfonate, and tosylate.

Protection for 1,2 and 1,3-diols

The protection for 1,2 and 1,3-diols group includes: cyclic acetals andketals, cyclic ortho esters, and silyl derivatives.

Cyclic acetals and ketals include: methylene, ethylidene,1-t-butylethylidene, 1-phenylethylidene, (4-methoxyphenyl)ethylidene,2,2,2-trichloroethylidene, acetonide(isopropylidene), cyclopentylidene,cyclohexylidene, cycloheptylidene, benzylidene, p-methoxybenzylidene,2,4-dimethoxybenzylidene, 3,4-dimethoxybenzylidene, and2-nitrobenzylidene.

Cyclic ortho esters include: methoxymethylene, ethoxymethylene,dimethoxy-methylene, 1-methoxyethylidene, 1-ethoxyethylidine,1,2-dimethoxyethylidene, α-methoxybenzylidene,1-(N,N-dimethylamino)ethylidene derivative,α-(N,N-dimethylamino)benzylidene derivative, and 2-oxacyclopentylidene.

Protection for the Carboxyl Group

Ester protecting groups include: esters, substituted methyl esters,2-substituted ethyl esters, substituted benzyl esters, silyl esters,activated esters, miscellaneous derivatives, and stannyl esters.

Substituted methyl esters include: 9-fluorenylmethyl, methoxymethyl,methylthiomethyl, tetrahydropyranyl, tetrahydrofuranyl,methoxyethoxymethyl, 2-(trimethylsilyl)ethoxy-methyl, benzyloxymethyl,phenacyl, p-bromophenacyl, α-methylphenacyl, p-methoxyphenacyl,carboxamidomethyl, and N-phthalimidomethyl.

2-Substituted ethyl esters include: 2,2,2-trichloroethyl, 2-haloethyl,1-chloroalkyl, 2-(trimethylsily)ethyl, 2-methylthioethyl,1,3-dithianyl-2-methyl, 2(p-nitrophenylsulfenyl)-ethyl,2-(p-toluenesulfonyl)ethyl, 2-(2′-pyridyl)ethyl,2-(diphenylphosphino)ethyl, 1-methyl-1-phenylethyl, t-butyl,cyclopentyl, cyclohexyl, allyl, 3-buten-1-yl,4-(trimethylsily)-2-buten-1-yl, cinnamyl, α-methylcinnamyl, phenyl,p-(methylmercaptoyphenyl, and benzyl.

Substituted benzyl esters include: triphenylmethyl, diphenylmethyl,bis(o-nitrophenyl)methyl, 9-anthrylmethyl, 2-(9,10-dioxo)anthrylmethyl,5-dibenzo-suberyl, 1-pyrenylmethyl,2-(trifluoromethyl)-6-chromylmethyl,2,4,6-trimethylbenzyl, p-bromobenzyl, o-nitrobenzyl, p-nitrobenzyl,p-methoxybenzyl, 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl,4-sulfobenzyl, piperonyl, and 4-P-benzyl.

Silyl esters include: trimethylsilyl, triethylsilyl,t-butyldimethylsilyl, i-propyldimethylsilyl, phenyldimethylsilyl, anddi-t-butylmethylsilyl.

Miscellaneous derivatives includes: oxazoles, 2-alkyl-1,3-oxazolines,4-alkyl-5oxo-1,3-oxazolidines, 5-alkyl-4-oxo-1,3-dioxolanes, orthoesters, phenyl group, and pentaaminocobalt(III) complex.

Examples of stannyl esters include: triethylstannyl andtri-n-butylstannyl.

Amides and Hydrazides

Amides include: N,N-dimethyl, pyrrolidinyl, piperidinyl,5,6-dihydrophenanthridinyl, o-nitroanilides, N-7-nitroindolyl,N-8-nitro-1,2,3,4-tetrahydroquinolyl, and p-P-benzenesulfonamides.Hydrazides include: N-phenyl, N,N′-diisopropyl and other dialkylhydrazides.

Protection for the Amino Group

Carbamates include: carbamates, substituted ethyl, assisted cleavage,photolytic cleavage, urea-type derivatives, and miscellaneouscarbamates.

Carbamates include: methyl and ethyl, 9-fluorenylmethyl,9-(2-sulfo)fluorenylmethyl, 9-(2,7-dibromo)fluorenylmethyl,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydro-thioxanthyl)]methyl,and 4-methoxyphenacyl.

Substituted ethyl protective groups include: 2,2,2-trichloroethyl,2-trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl,1,1-dimethyl-2-haloethyl, 1,1dimethyl-2,2-dibromoethyl,1,1-dimethyl-2,2,2-trichloroethyl, 1-methyl-1-(4-biphenylyl)ethyl,1-(3,5-di-t-butylphenyl)-1-methylethyl, 2-(2′-and 4′-pyridyl)ethyl,2-(N,N-icyclohexylcarboxamido)-ethyl, t-butyl, 1-adamantyl, vinyl,allyl, 1-isopropylallyl, connamyl, 4-nitrocinnamyl, quinolyl,N-hydroxypiperidinyl, alkyldithio, benzyl, p-methoxybenzyl,p-nitrobenzyl, p-bromobenzyl, p-chlorobenzyl, 2,4dichlorobenzyl,4-methylsulfinylbenzyl, 9-anthrylmethyl, and diphenylmethyl.

Protection via assisted cleavage includes: 2-methylthioethyl,2-methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl,[2-(1,3-dithianyl)]methyl, 4-methylthiophenyl, 2,4-dimethyl-thiophenyl,2-phosphonioethyl, 2-triphenylphosphonioisopropyl,1,1-dimethyl-2cyanoethyl, m-chloro-p-acyloxybenzyl,p-(dihydroxyboryl)benzyl, 5-benzisoxazolyl-methyl, and2-(trifluoromethyl)-6-chromonylmethyl.

Photolytic cleavage methods use groups such as: m-nitrophenyl,3,5-dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, andphenyl(o-nitrophenyl)methyl.

Examples of urea-type derivatives include: phenothiazinyl-(10)-carbonylderivative, N′-p-toluenesulfonylaminocarbonyl, andN′-phenylaminothiocarbonyl.

In addition to the above, miscellaneous carbamates include: t-amyl,S-benzyl thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl,cyclopentyl, cyclopropylmethyl, p-decyloxy-benzyl, diisopropylmethyl,2,2-dimethoxycarbonylvinyl, o-(N,N-dimethyl-carboxamido)-benzyl,1,1-dimethyl-3(N,N-dimethylcarboxamido)propyl, 1,1-dimethyl-propynyl,di(2-pyridyl)methyl, 2-furanylmethyl, 2-iodoethyl, isobomyl, isobutyl,isonicotinyl, p(p′-methoxyphenyl-azo)benzyl, 1-methylcyclobutyl,1-methylcyclohexyl, 1-methyl-1-cyclopropyl-methyl,1-methyl-(3,5-dimethoxyphenyl)ethyl, 1-methyl-1(p-henylazophenyl)-ethyl,1-methyl-1-phenylethyl, 1-methyl-1-(4-pyridyl)ethyl, phenyl,p-(phenylazo)benzyl, 2,4,6-tri-t-butylphenyl,4-(trimethylammonium)benzyl, and 2,4,6-trimethylbenzyl.

Amides

Amides includes: N-formyl, N-acetyl, N-chloroacetyl, N-trichloroacetyl,N-trifluoroacetyl, N-phenylacetyl, N-3-phenylpropionyl, N-picolinoyl,N-3-pyridyl-carboxamide, N-benzoylphenylalanyl derivative, N-benzoyl,and N-p-phenylbenzoyl.

Assisted cleavage groups include: N-o-nitrophenylacetyl,N-o-nitrophenoxyacetyl, N-acetoacetyl,(N′-dithiobenzyloxycarbonylamino)acetyl, N-3-(p-hydroxphenyl)propionyl,N-3-(o-nitrophenyl)propionyl, N-2-methyl-2-(o-nitrophenoxy)propionyl,N-2-methyl-2-(o-phenylazophenoxy)propionyl, N-4-chlorobutyryl,N-3-methyl-3-nitrobutyryl, N-o-nitrocinnamoyl, N-acetylmethioninederivative, N-o-nitrobenzoyl, N-o-(benzoyloxymethyl)benzoyl, and4,5-diphenyl-3-oxazolin-2-one.

Cyclic imide derivatives include: N-phthalimide, N-dithiasuccinoyl,N-2,3-diphenyl-maleoyl, N-2,5-dimethylpyrrolyl,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct, 5-substituted1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, and 1-substituted3,5-dinitro-4-pyridonyl.

Special—NH Protective Groups

Protective groups for —NH include: N-alkyl and N-aryl amines, iminederivatives, enamine derivatives, and N-hetero atom derivatives (such asN-metal, N—N, N—P, N—Si, and N—S), N-sulfenyl, and N-sulfonyl.

N-alkyl and N-aryl amines include: N-methyl, N-allyl,N-[2-(trimethylsilyl)ethoxyl]-methyl, N-3-acetoxypropyl,N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), quaternary ammoniumsalts, N-benzyl, N-di(4-methoxyphenyl)methyl, N-5-dibenzosuberyl,N-triphenylmethyl, N-(4-methoxyphenyl)diphenylmethyl,N-9-phenylfluorenyl, N-2,7-dichloro-9-fluorenylmethylene,N-ferrocenylmethyl, and N-2-picolylamine N′-oxide.

Imine derivatives include: N-1,1-dimethylthiomethylene, N-benzylidene,N-p-methoxybenzylidene, N-diphenylmethylene,N-[(2-pyridyl)mesityl]methylene, N(N′,N′-dimethylaminomethylene),N,N′-isopropylidene, N-p-nitrobenzylidene, N-salicylidene,N-5-chlorosalicylidene, N-(5-chloro-2-hydroxyphenyl)phenyl-methylene,and N-cyclohexylidene.

An example of an enamine derivative isN-(5,5-dimethyl-3-oxo-1-cyclohexenyl).

N-metal derivatives include: N-borane derivatives, N-diphenylborinicacid derivative, N-[phenyl(pentacarbonylchromium- or-tungsten)]carbenyl, and N-copper or N-zinc chelate. Examples of N—Nderivatives include: N-nitro, N-nitroso, and N-oxide. Examples of N—Pderivatives include: N-diphenylphosphinyl, N-dimethylthiophosphinyl,N-diphenylthiophosphinyl, N-dialkyl phosphoryl, N-dibenzyl phosphoryl,and N-diphenyl phosphoryl, Examples of N-sulfenyl derivatives include:N-benzenesulfenyl, N-o-nitrobenzenesulfenyl,N-2,4-dinitrobenzenesulfenyl, N-pentachlorobenzenesulfenyl,N-2-nitro-4-methoxy-benzenesulfenyl, N-triphenylmethylsulfenyl, andN-3-nitropyridinesulfenyl. N-sulfonyl derivatives include:N-p-toluenesulfonyl, N-benzenesulfonyl,N-2,3,6-trimethyl-4-methoxybenzenesulfonyl,N-2,4,6-trimethoxybenzenesulfonyl,N-2,6-dimethyl-4-methoxy-benzenesulfonyl, N-pentamethylbenzenesulfonyl,N-2,3,5,6-tetramethyl-4-methoxybenzene-sulfonyl,N-4-methoxybenzenesulfonyl, N-2,4,6-trimethylbenzenesulfonyl,N-2,6-dimethoxy-4-methylbenzenesulfonyl,N-2,2,5,7,8-pentamethylchroman-6-sulfonyl, N-methanesulfonyl,N-β-trimethylsilylethanesulfonyl, N-9-anthracenesulfonyl,N-4-(4′,8′-dimethoxynaphthylmethyl)-benzenesulfonyl, N-benzylsulfonyl,N-trifluoromethylsulfonyl, and N-phenacylsulfonyl.

Disclosed compounds which are masked or protected may be prodrugs,compounds metabolized or otherwise transformed in vivo to yield adisclosed compound, e.g., transiently during metabolism. Thistransformation may be a hydrolysis or oxidation which results fromcontact with a bodily fluid such as blood, or the action of acids, orliver, gastrointestinal, or other enzymes.

Some of these masked or protected compounds are pharmaceuticallyacceptable; others will be useful as intermediates. Syntheticintermediates and processes disclosed herein, and minor modificationsthereof, are also within the scope of the invention.

The compounds of formulas I and II can be prepared by techniques andprocedures readily available to one of ordinary skill in the art, forexample by following the procedures as set forth in the followingSchemes or analogous variants thereof. These synthetic strategies arefurther exemplified in Examples 1–6 below. These Schemes are notintended to limit the scope of the invention in any way.

As used herein, the following terms have the meanings indicated: “LiBH₄”refers to lithium borohydride; “TMSCI” refers to trimethylsilylchloride; “TBDPSCI” refers to tert-butyldiphenylsilyl chloride; “sBuLi”refers to sec-butyllithium; “TBAF” refers to tetrabutylammoniumfluoride; “HOAc” refers to acetic acid; “KMnO₄” refers to potassiumpermanganate; “LiHMDS” refers to lithium1,1,1,3,3,3-hexamethyl-disilazane. All other terms and substituents,unless otherwise indicated, are previously defined. The reagents andstarting materials are readily available to one of ordinary skill in theart. Scheme I provides a synthesis of the compound of structure (2).

In Scheme I, step a, the compound of structure (1), which is2,3,4-trifluorobenzoic acid, is reduced with in situ prepared boraneunder conditions described in Angew. Chem. Int. Ed. (1989), 28, 218 toprovide the corresponding alcohol.

In Scheme I, step b, the alcohol is protected with a suitable hydroxylprotecting group, such as tert-butyldiphenylsilyl chloride. A suitablehydroxyl protecting group will be stable to basic conditions.

In Scheme I, step c, directed metallation of the protected alcoholprovides the anion. In step d, the resulting anion is quenched withcarbon dioxide to provide the monoacid.

In Scheme I, steps e and f, the protected monoacid is deprotected andoxidized under conditions well known in the art to provide thesymmetrical diacid (2) which is 4,5,6-trifluoro-isophthalic acid.

These synthetic strategies are further exemplified in Example 1, stepsa–d.

Scheme II provides a synthesis of the compound of structure (4).

In Scheme II, step a, the compound of structure (3), which is2,4-difluorobenzoic acid, is reduced with in situ prepared borane underconditions described in Angew. Chem. Int. Ed. (1989), 28, 218 to providethe corresponding alcohol.

In Scheme II, step b, the alcohol is protected with a suitable hydroxylprotecting group, such as tert-butyldiphenylsilyl chloride. A suitablehydroxyl protecting group will be stable to basic conditions.

In Scheme II, step c, directed metallation of the protected alcoholprovides the anion. In step d, the resulting anion is quenched with asilylating agent, such as trimethylsilyl chloride to provide themonoacid.

In Scheme II, step e, directed metallation of the protected alcoholprovides the anion. In step f, the resulting anion is quenched withcarbon dioxide to provide the monoacid.

In Scheme II, steps g and h, the protected monoacid is deprotected andoxidized under conditions well known in the art to provide thesymmetrical diacid (4) which is 4,6-difluoroisophthalic acid.

Scheme III provides a synthesis of the compounds of formula I, whichincludes formulas Ia and Ib, and formula II.

In Scheme III, step A, a suitable aniline (5), such as 4-iodo-2methylaniline, 4-iodo-2-chloroaniline, or 4-iodo-2-fluoroaniline iscoupled with a symmetrical diacid (6) to provide the compound ofstructure (7). Examples of suitable diacids (6) include, but are notlimited to, compound (2) as shown in Scheme I, compound (4) as shown inScheme II and 4-fluoro-isophthalic acid which can be prepared by one ofordinary skill in the art following generally the procedure disclosed byChuprina. G. N. et al., Uzh. Vses. Khim. O-va, 19(5), 598–9 (1974). Itis within the skill of one of ordinary art to identify additionaldiacids (6) useful in the preparation of compounds of the presentinvention. For example, compound (6) and compound (5), in separateflasks, are each suspended in a suitable organic solvent, such astetrahydrofuran, at −78° C. under nitrogen. Each suspension is treatedwith an excess of a suitable base, such as 2 equivalents of lithium1,1,1,3,3,3-hexamethyl-disilazane or lithium amide. After both solutionsare stirred for about 30 minutes at −78° C., the diacid solution wastransferred by cannula into the aniline solution and allowed to warm toroom temperature. After stirring from about 4 to 12 hours, the mixturewas precipitated with a suitable solution, such as a saturated HCldiethyl ether solution or combined with 1N HCl and extracted with ethylacetate. The resulting precipitate was filtered and concentrated undervacuum to provide the anthranilic diacid (7).

In Scheme III, step B, the acidic groups of the diacid (7) aredifferentiated by protection using suitable aldehyde, such as formalinor paraformaldehyde when R1 is methyl; or by using methyl bromide andcesium fluoride when R1 is a halogen, such as chloride or fluoride. Forexample, the diacid (7), a suitable aldehyde, such as paraformaldehyde,and a suitable acid, such as para-toluenesulfonic acid monohydrate werecombined in a suitable solvent, such as dichloromethane. In aroundbottom flask attached with a Dean-Stark apparatus, the solution isallowed to reflux for about 3 hours. The resulting solution isconcentrated and the residue is suspended in a suitable solvent, such asmethanol. The aldehyde is filtered off, the filtrate is collected andconcentrated under vacuum to provide the free acid (8).

In Scheme III, step C, the free acid (8) is activated, such as by theaddition of trifluoroacetic acid pentafluorophenyl ester. For example,to a suspension of the free acid (8) in a suitable solvent, such as inN,N-dimethylformamide is added trifluoroacetic acid pentafluorophenylester and a suitable base, such as pyridine. The reaction mixture isstirred for about 4 hours, diluted with a suitable solvent, such asethyl acetate, and washed with a series of solutions, such as 3 timeswith 1.0 M HCl solution, 3 times with 5% aqueous NaHCO₃ solution, 2times with water and once with saturated brine solution. The organicextracts are combined, dried over sodium sulfate, filtered andconcentrated under vacuum to provide the activated free ester (9).

In Scheme III, step D, amines are added to the free ester (9) to providethe amide or the ester, which is a compound of formula II. For example,the free ester (9) is suspended in a suitable solvent, such astetrahydrofuran. To the resulting suspension, a suitable alcohol, suchas methanol, or suitable amines, such as methylamine hydrochloride and asuitable tertiary amine base, such as triethylamine andN,N-diisopropylethylamine are added. After stirring from about 12 to 17hours, the reaction mixture was diluted with a suitable solvent, such asethyl acetate and washed using a series of solutions, such as 2 timeswith water and 2 times with saturated brine solution. The organicextracts are combined, dried over sodium sulfate, filtered andconcentrated under vacuum to provide the amide of formula II.

Examples of X may be derived by one of ordinary skill in the art fromcommercially available reagents that include, but are not limited to,the following:

trans-2-aminocyclohexanol 2-amino-5-mercapto-1,3,4- hydrochloridethiadiazole 2-amino-1,3,4-thiadiazole 2-amino-5-methyl-1,3,4-thiadiazole3-amino-1-phenyl-2-pyrazolin-5-one 2-amino-5-ethyl-1,3,4-thiadiazole5-amino-3-methylisoxazole 2-amino-6-methoxybenzothiazole3-amino-5-methylisoxazole 2-amino-6-ethoxybenzothiazole5-amino-3-phenyl-1,2,4-thiadiazole 2-amino-6-methylbenzothiazole2-(2-aminoethyl)-1-methylpyrrolidine 2-amino-4-methylbenzothiazole2-(aminomethyl)-1-ethylpyrrolidine 4-aminobenzo-2,1,3-thiadiazole1-(2-aminoethyl)pyrrolidine 4-amino-6-chloro-2- methylmercaptopyrimidinepseudothiohydantoin 2-aminopyrimidine 1-(3-aminopropyl)-2-pyrrolidinone2-amino-4,6-dihydroxypyrimidine furfurylamine 4-aminopyrimidine1-aminomethyl-1-cyclohexanol aminopyrazine hydrochloride histamine4-morpholinoaniline 3-amino-1,2,4-triazole 4-(2-aminoethyl)morpholine3-amino-5-mercapto-1,2,4-triazole n-(3-aminopropyl)morpholine3-amino-5-methylthio-1,2,4-triazole 5-amino-2-chloropyridine3-aminopyrazole 5-amino-2-methoxypyridine 3-amino-4-carbethoxypyrazole2-aminopyridine 2-amino-2-thiazoline 2-aminopyridine 2-aminothiazole2-(aminomethyl)pyridine 2-amino-4-methylthiazole2-(2-aminoethyl)pyridine ethyl 2-amino-4-thiazoleacetate 3-aminopyridined-cycloserine 3-(aminomethyl)pyridine tetrahydrofurfurylamine4-aminopyridine thiophene-2-methylamine 4-(aminomethyl)pyridine2-aminopurine 3-amino-1,2,4-triazine 2-aminobenzimidazole1-(2-aminoethyl)piperidine 5-methoxytryptamine 3,4-ethylenedioxyaniline6-methoxytryptamine 2-aminophenethyl alcohol 6-aminoindazoleN,N-dimethyl-p-phenylenediamine 8-azaadenineN,N-diethyl-1,4-phenylenediamine 2-aminobenzothiazole2-aminobenzenesulfonamide 2-(2-aminoethoxy)ethanol sulfanilamide2-(3,4-dimethoxyphenyl)ethylamine 2-amino-1-methoxypropane3-isopropoxypropylamine dl-2-amino-1-propanol methyl 3-aminothiophene-2-4-hydroxypiperidine carboxylate n-(3-aminopropyl)imidazole4-piperidineethanol 3-aminopyrazine-2-carboxylic 1-methyl-4- acid methylester (methylamino)piperidine 5-amino-1-ethylpyrazoleN-methyl-p-anisidine 3-amino-5-hydroxypyrazole methylaminoacetaldehydedimethylacetal 2-amino-5-(ethylthio)-1,3,4- (S)-(+)-2-(methoxymethyl)thiadiazole pyrrolidine dl-cycloserine 1-methylpiperazinedihydrochloride 3-amino-5-methylpyrazole 3-hydroxypiperidinehydrochloride 4-chloro-n-methylaniline dl-nornicotine2-(methylamino)ethanol 4-hydroxypiperidine hydrochlorideN,N′-bis(2-hydroxyethyl) 4-(1-pyrrolidinyl)piperidine ethylenediaminediethanolamine N-ethylpiperazine 2-(butylamino)ethanethiol d-prolinolthiazolidine thialdine I-prolinol (R)-3-hydroxypyrrolidine3-pyrrolidinol (R)-(−)-3-pyrrolidinol hydrochlorideN-omega-methyltryptamine (R)-(+)-3-hydroxypiperidine hydrochloridepiperazine (S)-3-hydroxypyrrolidine 1-formylpiperazine thialdine1-methylpiperazine 1-benzylpiperazine N-(2-hydroxyethyl)piperazinemorpholine thiomorpholine 2-piperidinemethanol 2-piperidineethanol3-piperidinemethanol

In Scheme II, step E, the acid of the compound of formula II isdeprotected under acidic conditions using a polymer bound glycol as aquench reagent to provide the compound of formula Ia. For example, to asuspension of formula II in a suitable solvent, such as tetrahydrofuran,is added a suitable quench agent, such as polymer bound glycerol, and asuitable acidic solution, such as about 10 mL of 1.0 M hydrochloric acidsolution. After stirring for about 48 hours at room temperature, theresin is filtered off and the filtrate is transferred to a separatoryfunnel and partitioned with a suitable solvent, such as ethyl acetate.The organics are washed using a series of solutions, such as twice with1.0 M HCl and twice with saturated brine solution. The organic extractsare collected, dried over sodium sulfate, filtered and concentratedunder vacuum to provide the compound of formula Ia.

In Scheme III, step F, the deprotected acid of formula Ia is activated,such as by the addition of trifluoroacetic acid pentafluorophenyl esterand reacted with an appropriately substituted hydroxyl amine, to allowthe formation of the hydroxamate, which is the compound of formula 1b.For example, to a suspension of formula 1a in a suitable solvent, suchas in N,N-dimethylformamide is added trifluoroacetic acidpentafluorophenyl ester and a suitable base, such as pyridine. Thereaction mixture is stirred for about 17 hours, diluted with a suitablesolvent, such as ethyl acetate, and washed using a series of solutions,such as 3 times with 1.0 M HCl solution, 3 times with 5% aqueous NaHCO₃solution, 2 times with water and once with saturated brine solution. Theorganic extracts are combined, dried over sodium sulfate, filtered andconcentrated under vacuum to provide the activated free ester (10).

In Scheme III, step G, amines are added to the free ester (10) toprovide the amide, which is a compound of formula Ib. For example, thefree ester (10) is suspended in a suitable solvent, such astetrahydrofuran. To the resulting suspension, a suitable alcohol, suchas methanol, or suitable amines, such as methylamine hydrochloride andcyclopropylmethylamine hydrochloride, and a suitable tertiary aminebase, such as triethylamine and N,N-diisopropylethylamine are added.After stirring from about 12 to 17 hours, the reaction mixture waspartitioned between a suitable solvent, such as ethyl acetate and asuitable acid, such as 1.0 M HCl solution. The organic layer was washedusing a series of solutions, such as 2 times with water and 2 times withsaturated brine solution. The organic extracts are combined, dried overmagnesium sulfate, filtered and concentrated under vacuum to provide thehydroxamate of formula Ib.

One aspect of the invention features the disclosed compounds shown informulas I and II. Preferred compounds of formulas I or II are those inwhich R₁ is C₁₋₈ alkyl or halo, preferably C₁₋₈ alkyl, more preferablyfluoro, chloro, or methyl, and most preferably methyl; R3 and R4 areeach independently hydrogen or halo, preferably fluoro; A is hydroxy orNR₆OR₇; X is NR₁₃R₁₂ or NR₁₄; and R₁₂ and R₁₃ are each independently[(CH₂)_(n)Y(CH₂)_(m)]_(q)CH₃, (C₁₋₆ alkyl)phenyl,—[(CH₂)_(n)Y(CH₂)_(m)]_(q)phenyl, or (C₁₋₆ alkyl)C₂₋₆ heterocyclicradical.

Also preferred are compounds of formula I or formula II in whichheterocyclic radicals include heteroaryls such as substituted orunsubstituted radicals of pyrrole, furan, pyran, thiophene, pyrazole,imidazole, triazole, tetrazole, indole, isoxazole, indazole, pyridine,pyrazine, oxazole, thiazole, oxadiazole, oxathiadiazole; heterocyclesalso include heteroalkyls such as substituted and unsubstituted radicalsof morpholine, piperidine, piperazine, tetrahydrofuran, tetrahydropyran,pyrrolidone, imidazoline, and tetrahydrothiophene.

Table I and Table II provide examples of preferred compounds of thepresent invention.

TABLE I

—A —R₁ —X —OH -Me —NH₂ —OH -Me —NHMe —OH -Me —NMe₂ —OH -Me

—OH -Me

—OH -Me

—OH -Me —NCH₃nBu —OH -Me

—OH -Me —N[(CH₂)₂OCH₂CH₃]₂ —OH -Me

—OH -Me

—OH —Cl —NH₂ —OH —Cl —NHMe —OH —Cl —NMe₂ —OH —Cl

—OH —Cl

—OH —Cl

—OH —Cl —NCH₃nBu —OH —Cl

—OH —Cl —N[(CH₂)₂OCH₂CH₃]₂ —OH —Cl

—OH —Cl

—NHOCH₂cPr -Me —NH₂ —NHOCH₂cPr -Me —NHMe —NHOCH₂cPr -Me —NMe₂ —NHOCH₂cPr-Me

—NHOCH₂cPr -Me

—NHOCH₂cPr -Me

—NHOCH₂cPr -Me —NCH₃nBu —NHOCH₂cPr -Me

—NHOCH₂cPr -Me —N[(CH₂)₂OCH₂CH₃]₂ —NHOCH₂cPr -Me

—NHOCH₂cPr -Me

—NHOCH₂cPr —Cl —NH₂ —NHOCH₂cPr —Cl —NHMe —NHOCH₂cPr —Cl —NMe₂ —NHOCH₂cPr—Cl

—NHOCH₂cPr —Cl

—NHOCH₂cPr —Cl

—NHOCH₂cPr —Cl —NCH₃nBu —NHOCH₂cPr —Cl

—NHOCH₂cPr —Cl —N[(CH₂)₂OCH₂CH₃]₂ —NHOCH₂cPr —Cl

—NHOCH₂cPr —Cl

TABLE II

—R₁ —X -Me —NH₂ -Me —NHMe -Me —NMe₂ -Me

-Me

-Me

-Me —NCH₃nBu -Me

-Me —N[(CH₂)₂OCH₂CH₃]₂ -Me

-Me

—Cl —NH₂ —Cl —NHMe —Cl —NMe₂ —Cl

—Cl

—Cl

—Cl —NCH₃nBu —Cl

—Cl —N[(CH₂)₂OCH₂CH₃]₂ —Cl

—Cl

As used herein, the term “patient” refers to any warm-blooded animalsuch as, but not limited to, a human, horse, dog, guinea pig, or mouse.Preferably, the patient is human.

The term “treating” for purposes of the present invention refers toprophylaxis or prevention, amelioration or elimination of a namedcondition once the condition has been established.

According to one aspect of the invention, the compounds are MEKinhibitors. MEK inhibition assays include the in vitro cascade assay forinhibitors of MAP kinase pathway described at column 6, line 36 tocolumn 7, line 4 of U.S. Pat. No. 5,525,625 and the in vitro MEK assayat column 7, lines 4–27 of the same patent, the entire disclosure ofwhich is incorporated by reference (see also Examples 163–173 below).

Selective MEK 1 or MEK 2 inhibitors are those compounds which inhibitthe MEK 1 or MEK 2 enzymes, respectively, without substantiallyinhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase,EGF, and PDGF receptor kinases, and C-src. In general, a selective MEK 1or MEK 2 inhibitor has an IC₅₀ for MEK 1 or MEK 2 that is at leastone-fiftieth ( 1/50) that of its IC₅₀ for one of the above-named otherenzymes. Preferably, a selective inhibitor has an IC₅₀ that is at least1/100, more preferably 1/500, and even more preferably 1/1000, 1/5000,or less than that of its IC₅₀ or one or more of the above-named enzymes.

The disclosed compositions are useful as both prophylactic andtherapeutic treatments for diseases or conditions related to thehyperactivity of MEK, as well as diseases or conditions modulated by theMEK cascade. Examples include, but are not limited to, stroke, septicshock, heart failure, osteoarthritis, rheumatoid arthritis, organtransplant rejection, and a variety of tumors such as ovarian, lung,pancreatic, brain, prostatic, and colorectal.

The invention further relates to a method for treating proliferativediseases, such as cancer, restenosis, psoriasis, autoimmune disease, andatherosclerosis. Other aspects of the invention include methods fortreating MEK-related (including ras-related) cancers, whether solid orhematopoietic. Examples of cancers include brain, breast, lung, such asnon-small cell lung, ovarian, pancreatic, prostate, renal, colorectal,cervical, acute leukemia, and gastric cancer. Further aspects of theinvention include methods for treating or reducing the symptoms ofxenograft (cell(s), skin, limb, organ or bone marrow transplant)rejection, osteoarthritis, rheumatoid arthritis, cystic fibrosis,complications of diabetes (including diabetic retinopathy and diabeticnephropathy), hepatomegaly, cardiomegaly, stroke (such as acute focalischemic stroke and global cerebral ischemia), heart failure, septicshock, asthma, Alzheimer's disease, and chronic or neuropathic pain.Compounds of the invention are also useful as antiviral agents fortreating viral infections such as HIV, hepatitis (B) virus (HBV), humanpapilloma virus (HPV), cytomegalovirus (CMV), and Epstein-Barr virus(EBV). These methods include the step of administering to a patient inneed of such treatment, or suffering from such a disease or condition, apharmaceutically or therapeutically effective amount of a disclosedcompound or pharmaceutical composition thereof.

The term “chronic pain” for purposes of the present invention includes,but is not limited to, neuropathic pain, idiopathic pain, and painassociated with chronic alcoholism, vitamin deficiency, uremia, orhypothyroidism. Chronic pain is associated with numerous conditionsincluding, but not limited to, inflammation, arthritis, andpost-operative pain.

As used herein, the term “neuropathic pain” is associated with numerousconditions which include, but are not limited to, inflammation,postoperative pain, phantom limb pain, bum pain, gout, trigeminalneuralgia, acute herpetic and postherpetic pain, causalgia, diabeticneuropathy, plexus avulsion, neuroma, vasculitis, viral infection, crushinjury, constriction injury, tissue injury, limb amputation,post-operative pain, arthritis pain, and nerve injury between theperipheral nervous system and the central nervous system.

The invention also features methods of combination therapy, such as amethod for treating cancer, wherein the method further includesproviding radiation therapy or chemotherapy, for example, with mitoticinhibitors such as a taxane or a vinca alkaloid. Examples of mitoticinhibitors include paclitaxel, docetaxel, vincristine, vinblastine,vinorelbine, and vinflunine. Other therapeutic combinations include aMEK inhibitor of the invention and an anticancer agent such ascisplatin, 5-fluorouracil or 5-fluoro-2-4(1H,3H)-pyrimidinedione (5FU),flutamide, and gemcitabine.

The chemotherapy or radiation therapy may be administered before,concurrently, or after the administration of a disclosed compoundaccording to the needs of the patient.

Those skilled in the art will be able to determine, according to knownmethods, the appropriate therapeutically-effective amount or dosage of acompound of the present invention to administer to a patient, takinginto account factors such as age, weight, general health, the compoundadministered, the route of administration, the type of pain or conditionrequiring treatment, and the presence of other medications. In general,an effective amount or a therapeutically-effective amount will bebetween about 0.1 and about 1000 mg/kg per day, preferably between about1 and about 300 mg/kg body weight, and daily dosages will be betweenabout 10 and about 5000 mg for an adult subject of normal weight.Commercially available capsules or other formulations (such as liquidsand film-coated tablets) of 100 mg, 200 mg, 300 mg, or 400 mg can beadministered according to the disclosed methods.

The compounds of the present invention are preferably formulated priorto administration. Therefore, another aspect of the present invention isa pharmaceutical composition comprising a compound of formulas I or IIand a pharmaceutically acceptable carrier. In making the compositions ofthe present invention, the active ingredient, such as a compound offormula I or formula II, will usually be mixed with a carrier, ordiluted by a carrier or enclosed within a carrier. Dosage unit forms orpharmaceutical compositions include tablets, capsules, pills, powders,granules, aqueous and nonaqueous oral solutions and suspensions, andparenteral solutions packaged in containers adapted for subdivision intoindividual doses.

Dosage unit forms can be adapted for various methods of administration,including controlled release formulations, such as subcutaneousimplants. Administration methods include oral, rectal, parenteral(intravenous, intramuscular, subcutaneous), intracistemal, intravaginal,intraperitoneal, intravesical, local (drops, powders, ointments, gels,or cream), and by inhalation (a buccal or nasal spray).

Parenteral formulations include pharmaceutically acceptable aqueous ornonaqueous solutions, dispersion, suspensions, emulsions, and sterilepowders for the preparation thereof. Examples of carriers include water,ethanol, polyols (propylene glycol, polyethylene glycol), vegetableoils, and injectable organic esters such as ethyl oleate. Fluidity canbe maintained by the use of a coating such as lecithin, a surfactant, ormaintaining appropriate particle size. Carriers for solid dosage formsinclude (a) fillers or extenders, (b) binders, (c) humectants, (d)disintegrating agents, (e) solution retarders, (f) absorptionacccelerators, (g) adsorbants, (h) lubricants, (i) buffering agents, and(j) propellants.

Compositions may also contain adjuvants such as preserving, wetting,emulsifying, and dispensing agents; antimicrobial agents such asparabens, chlorobutanol, phenol, and sorbic acid; isotonic agents suchas a sugar or sodium chloride; absorption-prolonging agents such asaluminum monostearate and gelatin; and absorption-enhancing agents.

The following examples represent typical syntheses of the compounds offormula I and II as described generally above. These examples areillustrative only and are not intended to limit the invention in anyway. The reagants and starting materials are readily available to one ofordinary skill in the art. As used herein, the following terms have themeanings indicated: “g” refers to grams; “mg” refers to milligrams; “mL”refers to milliliters; “mmol” refer to millimoles; “° C.” refers todegrees Celsius; “APCI” refers to atmospheric pressure chemicalionization; and “THF” refers to tetrahydrofuran;

EXAMPLE 17,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid

-   Step a: To a suspension of (2,3,4-trifluoro-phenyl)-methanol    (prepared as in Angew. Chem. Int. Ed. (1989), 28,218) (8.7 g, 54    mmol) in 50 mL of dichloromethane was added tert-butyldiphenylsilyl    chloride (15.4 mL, 59 mmol) and imidazole (4.02 g, 59 mmol). After    17 hours, the reaction was poured into 100 mL of 1 M HCl solution    and extracted into dichloromethane. The organic layer was washed 2    times with 1 M HCl solution and 2 times with brine solution. The    organic phase was collected and dried over Na₂SO₄, filtered, and    concentrated in vacuo. The white, oily solid (18.8 g) was purified    by column silica chromatography eluting with 9:1 hexane:ethyl    acetate to afford 14.37 g (68.5%)    teft-butyl-diphenyl-(2,3,4-trifluoro-benzyloxy)-silane.-   Step b: To a suspension of    tert-butyl-diphenyl-(2,3,4-trifluoro-benzyloxy)-silane (7.42 g, 18.5    mmol) in freshly distilled tetrahydrofuran (50 mL) at −78° C. under    nitrogen is added 1.3 M sec-butyllithium in cyclohexane (18.5 mL,    24.0 mmol). The reaction was allowed to stir at −78° C. for 1 hour    and quenched with CO₂ gas (lecture bottle) directly into the    solution for 30. minutes and the reaction mixture was slowly brought    to room temperature. After 3 hours, the reaction was partitioned    between ethyl acetate and 1 M HCl solution and washed with brine    solution. The organic phase was collected and dried over Na₂SO₄,    filtered and concentrated in vacuo to afford 8.17 g (99.6%) of    5-(tert-butyldiphenyl-silanyloxymethyl)-2,3,4-trifluoro-benzoic acid    as a white, waxy solid.-   Step c: To a suspension of    5-(tert-butylphenyl-silanyloxymethyl)-2,3,4-trifluoro-benzoic acid    (8.17 g, 18.4 mmol) in freshly distilled THF (20 mL) was added a    solution of tetrabutylammonium fluoride (1.0 M in THF, 40.0 mL, 40.0    mmol). After stirring at room temperature for 2 hours the reaction    mixture was concentrated in vacuo and redissolved in ethyl acetate,    transferred to a separatory funnel and washed 3 times with 1M HCl    solution, 2 times with saturated brine solution. The organic layers    were collected, dried over Na₂SO₄, filtered and concentrated in    vacuo. To the resulting residue was added hexanes affording a white    solid, which was washed several times with hexanes, collected and    dried in vacuo affording 2,3,4-tifluoro-5-hydroxymethyl-benzoic acid    (1.98 g, 52.2%).-   Step d: To a refluxing suspension of    trifluoro-5-hydroxymethyl-benzoic acid (1.90 g, 9.22 mmol) in    acetone is added a solution of potassium permanganate (4.3 g, 27.7    mmol) in water (5 mL). After refluxing for 6 hours the reaction is    allowed to cool and an aqueous solution of NaHSO₃ (5 mL, 1.0 M) and    an aqueous solution of H₂SO₄ (5 mL, 1.0 M) is added which clears the    reaction solution. This mixture is transferred to a separatory    funnel and extracted several times with ethyl acetate. The organic    layers are collected, dried over Na₂SO₄, filtered and concentrated    in vacuo affording 4,5,6-trifluoro-isophthalic acid as a light    yellow solid (1.03 g, 50.7%).-   Step e: A suspension of 4,5,6-trifluoro-isophthalic acid (1.03 g,    4.68 mmol) in freshly distilled THF (20 mL) at −78° C. under    nitrogen is treated with 2.0 equivalents of freshly prepared 1 M    LiHMDS solution (HMDS, 2.07 mL, 9.83 mmol; n-butyllithium, 3.5 mL,    9.36 mmol) in THF. In a second flask is suspended    4-iodo-2-methylaniline (1.09 g, 4.68 mmol) in 20 mL of freshly    distilled THF, cooled to −78° C. under nitrogen and treated with 2.0    equiv. of freshly prepared 1M LiHMDS solution (HMDS, 2.07 mL, 9.83    mmol; n-butyllithium, 3.5 mL, 9.36 mmol) in THF. After both    solutions stirred for 30 minutes at −78° C., the benzoic acid    solution was cannula transferred into the aniline solution and    allowed to slowly warm to room temperature. After stirring for 4    hours, the reaction mixture was poured into 200 mL of a saturated    HCl diethyl ether solution affording a white precipitate. The solid    is filtered off and the remaining filtrate is collected and    concentrated in vacuo affording    4,5-fluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalic acid (1.55 g,    77%).-   Step f: A suspension of    4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalic acid (1.1    g, 2.54 mmol), paraformaldehyde (5.0 g), and para-toluenesulfonic    acid monohydrate (15.0 mg) in dichloromethane (250 mL) in a    roundbottom flask attached with a Dean-Stark apparatus is allowed to    reflux for 3 hours. The cooled solution is then concentrated and the    residue is redissolved in methanol and paraformaldehyde is filtered    off and filtrate is collected and concentrated in vacuo affording    7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylic    acid as a red solid (0.80 g, 70.0%).

EXAMPLE 27,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid methylamide

To a suspension of7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid (0.3 g, 0.67 mmol) in N,N-dimethylformamide (3 mL) is addedtrifluoroacetic acid pentafluorophenyl ester (0.127 mL, 0.74 mmol) andpyridine (0.60 mL, 0.74 mmol). After stirring for 4 hours the reactionmixture is diluted with ethyl acetate and washed 3 times with 1.0 M HClsolution, 3 times with 5% aqueous NaHCO₃ solution, 2 times with waterand once with saturated brine solution. The organic layers werecollected and dried over Na₂SO₄, filtered and concentrated in vacuoaffording7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid pentafluorophenylester as a dark orange oil (0.34 g, 83.1%). To asuspension of7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid pentafluorophenylester (0.33 g, 0.54 mmol) in freshly distilled THF(10 mL) is added methylamine hydrochloride (0.037 g, 0.54 mmol) andN,N-diisopropylethylamine (0.019 mL, 1.08 mmol). After stirring for 17hours, the reaction mixture was diluted with ethyl acetate and washed 2times with water and 2 times with saturated brine solution. The organiclayers were collected and dried over Na₂SO₄, filtered and concentratedin vacuo affording7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid methylamide as a yellow solid (0.19 g, 76.9%); mp 219–223° C.; ¹NMR(400 MHz; DMSO-d6) 8.40 (s, 1H), 8.06 (d, 1H, J=6.8), 7.74 (s, 1H), 7.50(d, 1H, J=8.4), 6.87 (d, 1H, J=8.0), 5.61 (s, 2H), 2.76 (d, 3H, J=4.4),2.25 (s, 3H); MS(APCI)m+1=459; Anal.calcd/found for C₁₇H₁₃F₂IN₂O₃ C45.00/45.39, H 3.01/3.16, N 6.05/5.88.

-   in vitro MEK assay: IC₅₀=6.6 μM

EXAMPLE 34,5-Difluoro-6-(-4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamicacid

To a suspension of7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid methylamide in THF (10 mL) is added polymer bound glycerol, (0.33g, 200–400 mesh) and 10 mL of 1.0 M HCl solution. After stirring for 48hours at room temperature, the resin is filtered off and the filtrate istransferred to a separatory funnel and partitioned with ethyl acetate.The organics are washed twice with 1.0 M HCl, twice with saturated brinesolution, collected, dried over Na₂SO₄, filtered and concentrated invacuo affording4,5-difluoro-6-(-4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamicacid (0.11 g, 76.2%); mp 254–259° C.; ¹H NMR (400 MHz; DMSO-d6) 9.31 (s,1H), 8.19 (s, 1H), 8.07 (d, 1H, J=7.2), 7.56 (s, 1H), 6.69 (t, 1H,J=5.6), 2.74 (d, 3H, J=4.4), 2.20 (s, 3H); MS (APCI)m+1=447;Anal.calcd/found for C₁₆H₁₃F₂IN₂O₃, C 43.07/43.26, H 2.94/3.07. N6.28/6.10.

-   in vitro MEK assay: IC₅₀=2.4 μM

EXAMPLE 4N¹-Cycloprolylmethoxy-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-N³-methyl-isophthalamide

To a suspension of4,5-difluoro-6-(-4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamicacid (0.13 g, 0.29 mmol) in N,N-dimethylformamide (5 mL) is addedtrifluoroacetic acid pentafluorophenyl ester (0.055 mL, 0.32 mmol) andpyridine (0.03 mL, 0.32 mmol). After stirring for 17 hours the reactionmixture is diluted with ethyl acetate and transferred to a separatoryfunnel, washed twice with 1.0 M HCl, twice with 5% aqueous NaHCO₃solution, 2 times with water and once with saturated brine solution. Theorganic layers were collected and dried over Na₂SO₄, filtered andconcentrated in vacuo affording4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamic acidpentafluorophenyl ester (0.08g, 45.2%). To a suspension of4,5-difluoro-6-(-4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamicacid pentafluorophenyl ester (0.08 g, 0.13 mmol) in freshly distilledTHF (3 mL) is added cyclopropylmethylamine hydrochloride (0.016 g, 0.13mmol), and N,N-diisopropylethylamine (0.07 mL, 0.39 mmol). Afterstirring at room temperature for 17 hours the reaction mixture waspartitioned between ethyl acetate and 1.0 M HCl solution. The organiclayer was washed twice with water, twice with saturated brine solution,dried over MgSO₄, filtered and concentrated in vacuo. Purification wasperformed by silica column chromatography in 2:1 ethyl acetate:hexanesaffordingN¹-cyclopropylmethoxy-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-N³-methyl-isophthalamide(0.033, 42.8%); mp 198–202° C.; ¹H NMR (400 MHz; DMSO-d6) 8.59 (s, 1H),8.01 (s, 1H), 7.44 (d, 1H, J=5.2), 7.32 (s, 1H), 7.18 (d, 1H, J=7.6),6.36 (m,1H), 3.34 (d, 2H, J=6.8), 2.56 (d, 3H, J=4.0), 1.99 (s, 3H),0.82 (m, 1H), 0.27 (d, 2H, J=8.4), 0.00 (m, 2H); MS(APCI)m+1=516;

-   Anal.calcd/found for C₂₀H₂₀F₂IN₃O₃ C 47.08/46.85, H 4.22/4.02 N    7.68/7.29.-   in vitro MEK assay: IC₅₀=1.1 μM

EXAMPLE 57,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid dimethylamide

Prepared in the manner of Example 4

-   ¹H NMR (400 MHz; CDCl₃) 7.96 (dd, 1H, J=2.2, 6.6), 7.65 (t, 1H,    J=1.5), 7.48 (dd, 1H, J=1.7, 8.3), 6.70 (d, 1H, J=8.3), 5.37 (s,    2H), 3.11 (s, 3H), 2.96 (s, 3H), 2.3 (s, 3H); MS(APCI)m+1=473;    Anal.calcd/found for C₁₈F₁₅F₂IN₂O₃ C 46.66/47.05, H 3.80/3.65, N    5.34/5.58.-   in vitro MEK assay: IC₅₀=5.8 μM

EXAMPLE 6N¹-Cyclopropylmethoxy-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-N³,N³-dimethyl-isophthalamide(27)

Prepared in the manner of Example 4

-   mp 78–80° C.; ¹H NMR (400 MHz; DMSO-d₆) 8.52 (s, 1H), 7.32 (s, 1H),    7.15 (m, 1H), 6.39 (m, 1H), 3.35 (d, 2H, J=6.8), 2.79 (s, 3H), 2.72    (s, 3H), 2.00 (s, 3H), 0.93 (m, 1H), 0.27 (d, 2H, J=8.0), 0.00 (m,    2H); MS(APCI)m+1=530.-   in vitro MEK assay: IC₅₀=4.3 μM

EXAMPLE 7

The following were prepared using parallel synthetic techniques in thefollowing manner:

Step A:

A solution of7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid pentafluorophenylester in a 2:1 mixture of THF toN,N-dimethylformamide (0.32M, 19.36 g) was prepared. In preweighed2-dram glass vials was added the corresponding amine (0.35 mmol) andthen the prepared7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid pentafluorophenylester solution (0.3 mmol). To each vial was addeda morpholine polystyrene resin (0.2 g), capped with Teflon coated capsand placed on an orbital shaker for 24 hours. The individual reactionswere then charged with polyamine polystyrene resin (0.2 g) andisocyanate polystyrene resin (0.1 g) and dichloromethane (2 mL) andallowed to shake for another 17 hours. The reactions were filtered andconcentrated in vacuo to afford the corresponding7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid amides. LC/MS was performed on a CPI 120SE (C18) column (4.6×50 mm,5 μm).

EXAMPLE 87,8-Difluoro-6-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₀H₁₇F₂IN₂O₄, MS (APCI)m+1=515-   in vitro MEK assay: 34% inhibition @ 1 μM

EXAMPLE 97,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-6-((S)-2-methoxymethyl-pyrrolidine-1-carbonyl)-1,2-dihydro-3,1-benzoxazin-4-one

C₂₂H₂₁F₂IN₂O₄, MS (APCI)m+1=543

-   in vitro MEK assay: 93% inhibition @ 1 μM

EXAMPLE 107,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (2-piperidin-1-yl-ethyl)-amide

-   C₂₃H₂₄F₂IN₃O₃, MS (APCI)m+1=556-   in vitro MEK assay: 44% inhibition @ 1 μM

EXAMPLE 117,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (2-pyrrolidin-1-yl-ethyl)-amide

-   C₂₂H₂₂F₂IN₃O₃, MS (APCI)m+1+542-   in vitro MEK assay: 33% inhibition @ 1 μM

EXAMPLE 127,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [3-(2-oxo-pyrrolidin-1-y)-propyl]-amide

-   C₂₃H₂₂F₂IN₃O₄, MS (APCI)m+1=570-   in vitro MEK assay: 21% inhibition @ 1 μM

EXAMPLE 137,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (1-hydroxy-cyclohexylmethyl)-amide

-   C₂₃H₂₃F₂IN₂O₄, MS (APCI)m+1=557-   in vitro MEK assay: 23% inhibition @ 1 μM

EXAMPLE 147,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (pyridin-2-ylmethyl)-amide

-   C₂₇H₂₄F₂IN₃O₃, MS (APCI)m+1=604

EXAMPLE 154-[7,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carbonyl]-piperazine-1-carbaldehyde

-   C₂₁H₁₈F₂IN₃O₄, MS (APCI)m+1=542-   in vitro MEK assay: 13% inhibition @ 1 μM

EXAMPLE 167,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid methyl-(1-methyl-piperidin-4-ylamide

-   C₂₃H₂₄F₂IN₃O₃, MS (APCI)m+1=556-   in vitro MEK assay: 0% inhibition @ 1 μM

EXAMPLE 177,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-6-(4-methyl-piperazine-1-carbonyl-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₁H₂₀F₂IN₃O₃, MS (APCI)m+1=528-   in vitro MEK assay: 78% inhibition @ 1 μM

EXAMPLE 187,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(2-hydroxy-ethoxy)-ethyl]-amide

-   C₂₀H₁₉F₂IN₂O₅, MS (APCI)m+1=533-   in vitro MEK assay: 43% inhibition @ 1 μM

EXAMPLE 197,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(1-methyl-pyrrolidin-2-yl)ethyl]-amide

-   C₂₃H₂₄F₂IN₃O₃, MS (APCI)m+1=556-   in vitro MEK assay: 28% inhibition @ 1 μM

EXAMPLE 207,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (2-hydroxy-ethyl)-methyl-amide

-   C₁₉H₁₇F₂IN₂O₄, MS (APCI)m+1=503-   in vitro MEK assay: 41% inhibition @ 1 μM

EXAMPLE 217,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid 1,3,4-thiadiazol-2-ylamide

-   C₁₈H₁₁F₂IN₄O₃S, MS (APCI)m+1=529-   in vitro MEK assay: 31% inhibition @ 1 μM

EXAMPLE 22

7,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (2-methoxy-1-methyl-ethyl)-amide

-   C₂₀H₁₉F₂IN₂O₄, MS (APCI)m+1=517-   in vitro MEK assay: 46% inhibition @ 1 μM

EXAMPLE 237,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (4-methyl-benzothiazol-2-yl)-amide

-   C₂₄H₁₆F₂IN₃O₃S, MS (APCI)m+1=592-   in vitro MEK assay: 26% inhibition @ 1 μM

EXAMPLE 24,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4dihydro-2H-3,1-benzoxazine-6-carboxylicacid (4-methyl-thiazol-2-yl)-amide

-   C₂₀H₁₄F₂IN₃O₃S, MS (APCI)m+1=542-   in vitro MEK assay: 20% inhibition @ 1 μM

EXAMPLE 257,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (5-ethylsulfanyl-1,3,4-thiadiazol-2-yl)-amide

-   C₂₀H₁₅F₂IN₄O₃S₂, MS (APCI)m+1=589-   in vitro MEK assay: 5% inhibition @ 1 μM

EXAMPLE 267,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (5-ethyl-1,3,4-thiadiazol-2-yl)-amide

-   C₂₀H₁₅F₂IN₄O₃S, MS (APCI)m+1=557-   in vitro MEK assay: 20% inhibition @ 1 μM

EXAMPLE 277,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (5-mercapto-1,3,4-thiadiazol-2-yl)-amide

-   C₁₈H₁₁F₂IN₄O₃S₂, MS (APCI)m+1=561-   in vitro MEK assay: 27% inhibition @ 1 μM

EXAMPLE 287,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (6-ethoxy-benzothiazol-2-yl)-amide

-   C₂₅H₁₈F₂IN₃O₄S, MS (APCI)m+1=622-   in vitro MEK assay: 9% inhibition @ 1 μM

EXAMPLE 297,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid benzothiazol-2-ylamide

-   C₂₃H₁₄F₂IN₃O₃S, MS (APCI)m+1=578-   in vitro MEK assay: 21% inhibition @ 1 μM

EXAMPLE 307,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(2-hydroxy-ethyl)-phenyl]-amide

-   C₂₄H₁₉F₂IN₂O₄, MS (APCI)m+1=565-   in vitro MEK assay: 55% inhibition @ 1 μM

EXAMPLE 317,8-Difluoro-1-(4-iodo-2-methyl-phenul)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid thiazol-2-ylamide

-   C₁₉H₁₂F₂IN₃O₃S, MS (APCI)m+1=528-   in vitro MEK assay: 32% inhibition @ 1 μM

EXAMPLE 327,8-Difluoro-6-[2-(2-hydroxy-ethyl)-piperidine-1-carbonyl]-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₃H₂₃F₂IN₂O₄, MS (APCI)m+1=557-   in vitro MEK assay: 76% inhibition @ 1 μM

EXAMPLE 337,8-Difluoro-6-(2-hydroxymethyl-piperidine-1-carbonyl)-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₂H₂₁F₂IN₂O₄, MS (APCI)m+1=543-   in vitro MEK assay: 79% inhibition @ 1 μM

EXAMPLE 343-{[7,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carbonyl]-amino}-1H-pyrazole-4-carboxylicacid ethyl ester

-   C₂₂H₁₇F₂IN₄O₅, MS (APCI)m+1=583-   in vitro MEK assay: 72% inhibition @ 1 μM

EXAMPLE 357,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (5-methylsulfanyl-1H-1,2,4-triazol-3-yl)-amide

-   C₁₉H₁₄F₂IN₅O₃S, MS (APCI)m+1=558-   in vitro MEK assay: 76% inhibition @ 1 μM

EXAMPLE 367,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (1H-pyrazol-3-yl)-amide

-   C₁₉H₁₃F₂IN₄0₃, MS (APCI)m+1=511-   in vitro MEK assay: 60% inhibition @ 1 μM

EXAMPLE 377,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid pyridin-3-ylamide

-   C₂₁H₁₄F₂IN₃O₃, MS (APCI)m+1=522-   in vitro MEK assay: 20% inhibition @ 1 μM

EXAMPLE 387,8-Difluoro-6-(3-hydroxy-piperidine-1-carbonyl)-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₁H₁₉F₂IN₂O₄, MS (APCI)m+1=529-   in vitro MEK assay: 48% inhibition @ 1 μM

EXAMPLE 397,8-Difluoro-6-(3-hydroxymethyl-piperidine-1-carbonyl)-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₂H₂₁F₂IN₂O₄, MS (APCI)m+1=543-   in vitro MEK assay: 71% inhibition @ 1 μM

EXAMPLE 407,8-Difluoro-6-(3-hydroxy-pyrrolidine-1-carbonyl)-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₀H₁₇F₂IN₂O₄, MS (APCI)m+1=515-   in vitro MEK assay: 42% inhibition @ 1 μM

EXAMPLE 417,8-Difluoro-1-(4-iodo-2-methyl-phenyl-6-(4-pyrrolidin-1-yl-piperidine-1-carbonyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₅H₂₆F₂IN₃O₃, MS (APCI)m+1=582-   in vitro MEK assay: 7% inhibition @ 1 μM

EXAMPLE 427,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (2-morpholin-4yl-ethyl)-amide

-   C₂₂H₂₂F₂IN₃O₄, MS (APCI)m+1=558-   in vitro MEK assay: 15% inhibition @ 1 μM

EXAMPLE 437,8-Difluoro-6-(4-hydroxy-piperidine-1-carbonyl)1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₁H₁₉F₂IN₂O₄, MS (APCI)m+1=529-   in vitro MEK assay: 52% inhibition @ 1 μM

EXAMPLE 447,8-Difluoro-6-[4-(2-hydroxy-ethyl)-piperidine-1-carbonyl]-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₃H₂₃F₂IN₂O₄, MS (APCI)m+1=557-   in vitro MEK assay: 69% inhibition @ 1 μM

EXAMPLE 457,8-Difluoro-1-(4-iodo-2-methyl-phenyl4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (6-chloro-pyridin-3-yl)-amide

-   C₂₁H₁₃ClF₂IN₃O₃, MS (APCI)m+1=556-   in vitro MEK assay: 62% inhibition @ 1 μM

EXAMPLE 467,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (6-methoxy-pyridin-3-yl)amide

-   C₂₂H₁₆F₂IN₃O₄, MS (APCI)m+1=552-   in vitro MEK assay: 43% inhibition @ 1 μM

EXAMPLE 477,8-Difluoro-1-(4-iodo-2-methyl-2-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(5-methoxy-1H-indol-3-yl)ethyl]-amide

-   C₂₇H22F₂IN₃O₄, MS (APCI)m+1=618-   in vitro MEK assay: 0% inhibition @ 1 μM

EXAMPLE 487,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(6-methoxy-1H-indol-3-yl)-ethyl]-amide

-   C₂₇H22F₂IN₃O₄, MS (APCI)m+1=618-   in vitro MEK assay: 12% inhibition @ 1 μM

EXAMPLE 497,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid bis-(2-hydroxy-ethyl)-amide

-   C₂₀H₁₉F₂IN₂O₅, MS (APCI)m+1=533-   in vitro MEK assay: 50% inhibition @ 1 μM

EXAMPLE 507,8-Difluoro-1-(4-iodo-2-methyl-phenyl)4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(1H-imidazol4-yl)-ethyl]-amide

-   C₂₁H₁₇F₂IN₄O₃, MS (APCI)m+1=539-   in vitro MEK assay: 44% inhibition @ 1 μM

EXAMPLE 517,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-6-(morpholine-4-carbonyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₀H₁₇F₂IN₂O₄, MS (APCI)m+1=515-   in vitro MEK assay: 72% inhibition @ 1 μM

EXAMPLE 527,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (3-imidazol-1-yl-propyl)amide

-   C₂₂H₁₉F₂IN₄O₃, MS (APCI)m+1 =553-   in vitro MEK assay: 35% inhibition @ 1 μM

EXAMPLE 537,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (4-dimethylamino-phenyl)amide

-   C₂₄H₂₀F₂IN₃O₃, MS (APCI)m+1=564-   in vitro MEK assay: 61% inhibition @ 1 μM

EXAMPLE 546-(4-Ethyl-piperazine-1-carbonyl)-7,8-difluoro-1-(4-iodo-2-methyl-phenyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₂H₂₂F₂IN₃O₃, MS (APCI)m+1=542-   in vitro MEK assay: 56% inhibition @ 1 μM

EXAMPLE 557,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid [2-(1H-indol-3-yl)ethyl ]-methyl-amide

-   C₂₇H₂₂F₂IN₃O₃, MS (APCI)m+1=602-   in vitro MEK assay: 41% inhibition @ 1 μM

EXAMPLE 567,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-6-(piperazine-1-carbonyl)-1,2-dihydro-3,1-benzoxazin-4-one

-   C₂₀H₁₈F₂IN₃O₃, MS (APCI)m+1=514-   in vitro MEK assay: 1% inhibition @ 1 μM

EXAMPLE 577,8-Difluoro-1-(4-iodo-2-methyl-phenyl)4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid (tetrahydro-furan-2-ylmethyl)-amide

-   C₂₁H₁₉F₂IN₂O₄, MS (APCI)m+1=529-   in vitro MEK assay: 35% inhibition @ 1 μM

EXAMPLE 587,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-6-(thiazolidine-3-carbonyl)-1,2-dihydro-3,1-benzoxzin-4-one

-   C₁₉H₁₅F₂IN₂O₃S, MS (APCI)m+1=517-   in vitro MEK assay: 76% inhibition @ 1 μM

EXAMPLE 597,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-6-(thiomorpholine-4-carbonyl)-1,2-dihydro-3,1-benzoxzin-4-one

-   C₂₀H₁₇F₂IN₂O₃S, MS (APCI)m+1=531-   in vitro MEK assay: 75% inhibition @ 1 μM

EXAMPLE 607,8-difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-3,1-benzoxazine-6-carboxylicacid ((S)-2-hydroxy-cyclohexyl)-amide

-   C₂₂H₂₁F₂IN₂O₄, MS (APCI)m+1=543-   in vitro MEK assay: 30% inhibition @ 1 μM    Step B:

To each of the 2 dram vials containing the7,8-Difluoro-1-(4-iodo-2-methyl-phenyl)-4-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazine-6-carboxylicacid amides was added THF (1 mL) and aqueous HCl (1.0 M, 1 mL) andglycerol polystyrene resin (0.2 g), capped with Teflon coated caps andallowed to shake on an orbital shaker at 50° C. for 5 days. Thereactions were filtered and washed with ethyl acetate (1.5 mL) andconcentrated in vacuo. HPLC purification was performed inacetonitrile/water (0.05%TFA) on a YM C30 (C18) column (100 mm ODS-A) toafford the corresponding isophthalamic acids. LC/MS was performed on aCPI 120SE (C18) column (4.6×50 mm, 5 μm).

EXAMPLE 613,4-Difluoro-5-[1-((R)-3-hydroxy-pyrrolidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₁₉H₁₇F₂IN₂O₄, MS (APCI)m+1=503-   in vitro MEK assay: 75% inhibition @ 1 μM

EXAMPLE 624,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-[3-(2-oxo-pyrrolidin-1-yl)-propyl]-isophthalamicacid

-   C₂₂H₂₂F₂IN₃O₄, MS (APCI)m+1=558-   in vitro MEK assay: 78% inhibition @ 1 μM

EXAMPLE 634,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(1-methyl-piperidin-4-yl)-isophthalamicacid

-   C₂₂H₂₄F₂IN₃O₃, MS (APCI)m+1=544-   in vitro MEK assay: 0% inhibition @ 1 μM

EXAMPLE 643,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(4-methyl-piperazin-1-yl)-methanoyl]-benzoicacid

-   C₂₀H₂₀F₂IN₃O₃, MS (APCI)m+1=516-   in vitro MEK assay: 57% inhibition @ 1 μM

EXAMPLE 654,5-Difluoro-N-[2-(2-hydroxy-ethoxy)-ethyl]-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₁₉H₁₉F₂IN₂O₅, MS (APCI)m+1=521-   in vitro MEK assay: 63% inhibition @ 1 μM p0 in vitro MEK assay:    IC₅₀=1.38 μM

EXAMPLE 664,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-[2-(1-methyl-pyrrolidin-2-yl)-ethyl]-isophthalamicacid

-   C₂₂H₂₄F₂IN₂O₃, MS (APCI)m+1=544-   in vitro MEK assay: 14% inhibition @ 1 μM

EXAMPLE 674,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-2-yl-ethyl)-isophthalamicacid

-   C₂₂H₁₈F₂IN₃O₃, MS (APCI)m+1=538-   in vitro MEK assay: 81% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=3.3 μM

EXAMPLE 68N-Butyl-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(2-mercapto-ethyl)-isophthalamicacid

-   C₂₁H₂₃F₂IN₂O₃S, MS (APCI)m+1=549-   in vitro MEK assay: 24% inhibition @ 1 μM

EXAMPLE 693,4-Difluoro-5-{1-[2-(2-hydroxy-ethyl)-piperidin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₂H₂₃F₂IN₂O₄, MS (APCI)m+1=545-   in vitro MEK assay: 78% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=2.2 μM

EXAMPLE 703,4-Difluoro-5-[1-(2-hydroxymethyl-piperidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₁H₂₁F₂IN₂O₄, MS (APCI)m+1=531-   in vitro MEK assay: 49% inhibition @ 1 μM

EXAMPLE 714,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-pyridin-3-ylmethyl-isophthalamicacid

-   C₂₁H₁₆F₂IN₃O₃, MS (APCI)m+1=524-   in vitro MEK assay: 86% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=1.45 μM

EXAMPLE 72N-(2,3-Dihydro-benzo[1,4]dioxin-6-yl)4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₃H₁₇F₂IN₂O₅, MS (APCI)m+1=567-   in vitro MEK assay: 65% inhibition@ 1 μM

EXAMPLE 733-({1-[5-Carboxy-2,3-difluoro-4-(4-iodo-2-methyl-phenylamino)-phenyl]-methanoyl}-amino)-1H-pyrazole-4-carboxylicacid ethyl ester

-   C₂₁H₁₇F₂IN₄O₅, MS (APCI)m+1=571-   in vitro MEK assay: 86% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=1.1 μM

EXAMPLE 744,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-pyridin-3-yl-isophthalamicacid

-   C₂₀H₁₄F₂IN₃O₃, MS (APCI)m+1=510-   in vitro MEK assay: 50% inhibition @ 1 μM

EXAMPLE 751-[3,4-Difluoro-5-[1-(3-hydroxy-piperidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-phenyl]-ethanone

-   C₂₀H₁₉F₂IN₂O₄, MS (APCI)m+1=517-   in vitro MEK assay: 92% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.465 μM

EXAMPLE 763,4-Difluoro-5-[1-(3-hydroxymethyl-piperidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₁H₂₁F₂IN₂O₄, MS (APCI)m+1=531-   in vitro MEK assay: 88% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.300 μM

EXAMPLE 773,4-Difluoro-5-[1-(3-hydroxy-pyrrolidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₁₉H₁₇F₂IN₂O₄, MS (APCI)m+1=503-   in vitro MEK assay: 83% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.880 μM

EXAMPLE 783,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(4-pyrrolidin-1-yl-piperidin-1-yl)-methanoyl]-benzoicacid

-   C₂₄H₂₆F₂IN₃O₃, MS (APCI)m+1=570-   in vitro MEK assay: 33% inhibition @ 1 μM

EXAMPLE 794,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-isophthalamicacid

-   C₂₁H₁₆F₂IN₃O₄, MS (APCI)m+1=546-   in vitro MEK assay: 54% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=1.5 μM

EXAMPLE 804,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-pyridin4-ylmethyl-isophthalamicacid

-   C₂₁H₁₆F₂IN₃O₃, MS (APCI)m+1=524-   in vitro MEK assay: 71% inhibition 1 μM-   in vitro MEK assay: IC₅₀=1.7 μM

EXAMPLE 813,4-Difluoro-5-[1-(4-hydroxy-piperidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₀H₁₉F₂IN₂O₄, MS (APCI)m+1=517-   in vitro MEK assay: 86% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=1.8 μM

EXAMPLE 824,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(4-morpholin-4-yl-phenyl)-isophthalamicacid

-   C₂₅H2F₂IN₃O₄, MS (APCI)m+1=594

EXAMPLE 833,4-Difluoro-5-{1-[4-(2-hydroxy-ethyl)-piperidin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₂H₂₃F₂IN₂O₄, MS (APCI)m+1=545-   in vitro MEK assay: 90% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.150 μM

EXAMPLE 84N-(2-Ethyl-2H-pyrazol-3-yl)-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₀H₁₇F₂IN₄O₃, MS (APCI)m+1=527-   in vitro MEK assay: 79% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.960 μM

EXAMPLE 85N-(6-Chloro-pyridin-3-yl)-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₀H₁₃F₂IN₃O₃, MS (APCI)m+1=544-   in vitro MEK assay: 90% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.970 μM

EXAMPLE 864,5-Difluoro-N-(1H-indazol-6-yl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₂H₁₅F₂IN₄O₃, MS (APCI)m+1=549-   in vitro MEK assay: 77% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=1.1 μM

EXAMPLE 874,5-Difluoro-N-(2-hydroxy-1-methyl-ethyl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₁₈H₁₇F₂IN₂O₄, MS (APCI)m+1 491-   in vitro MEK assay: 66% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=2.2 μM

EXAMPLE 884,5-Difluoro-N-[2-(1H-imidazol-4-yl)-ethyl]-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₀H₁₇F₂IN₄O₃, MS (APCI)m+1=527-   in vitro MEK assay: 60% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=1.4 μM

EXAMPLE 893,4-Difluoro-2-(4-iodo-2-methyl-1-phenylamino)-5-(1-morpholin-4-yl-methanoyl)-benzoicacid

-   C₁₉H₁₇F₂IN₂O₄, MS (APCI)m+1=503-   in vitro MEK assay: 89% inhibition @ 1 μM

EXAMPLE 904,5-Difluoro-N-(3-imidazol-1-yl-propyl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₁H₁₉F₂IN₄O₃, MS (APCI)m+1=541-   in vitro MEK assay: 59% inhibition @ 1 μM

EXAMPLE 914,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(3-morpholin4-yl-propyl)-isophthalamicacid

-   C₂₂H₂₄F₂IN₃O₄, MS (APCI)m+1=560-   in vitro MEK assay: 58% inhibition @ 1 μM

EXAMPLE 92N-(4-Dimethylamino-phenyl)-4,5-difluoro-6-(iodo-methyl-phenylamino)-isophthalamicacid

-   C₂₃H₂₀F₂IN₃O₃, MS (APCI)m+1=552-   in vitro MEK assay: 80% inhibition @ 1 μM

EXAMPLE 935-[1-(4-Ethyl-piperazin-1-yl)-methanoyl]-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₁H22F₂IN₃O₃, MS (APCI)m+1=530-   in vitro MEK assay: 70% inhibition @ 1 μM

EXAMPLE 944,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(4-methoxy-phenyl)-N-methyl-isophthalamicacid

-   C₂₃H₁₉F₂IN₂O₄, MS (APCI)m+1=553-   in vitro MEK assay: 92% inhibition @ 1 μM

EXAMPLE 954,5-Difluoro-N-[2-(1H-indol-3-yl)-ethyl]-6-(4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamicacid

-   C₂₆H₂₂F₂IN₃O₃, MS (APCI)m+1=590-   in vitro MEK assay: 77% inhibition @ 1 μM

EXAMPLE 964,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-phenyl)-isophthalamicacid

-   C₂₁H₁₆F₂IN₃O₄S, MS (APCI)m+1=588-   in vitro MEK assay: 85% inhibition @ 1 μM

EXAMPLE 974,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(tetrahydro-furan-2-ylmethyl)-isophthalamicacid

-   C₂₀H₁₉F₂IN₂O₄, MS (APCI)m+1=517-   in vitro MEK assay: 81% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.150 M

EXAMPLE 983,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(1-thiazolidin-3-yl-methanoyl)-benzoicacid

-   C₁₈H₁₅F₂IN₂O₃S, MS (APCI)m+1=505-   in vitro MEK assay: 86% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.087 μM

EXAMPLE 993,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(1-thiomorpholin-4-yl-methanoyl-benzoicacid

-   C₁₉H17F₂IN₂O₃S, MS (APCI)m+1=519-   in vitro MEK assay: 82% inhibition @ 1 μM-   in vitro MEK assay: IC₅₀=0.150 μM

EXAMPLE 1004,5-Difluoro-N-((S)-2-hydroxy-cyclohexyl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₁H₂₁F₂IN₂O₄, MS (APCI)m+1=531

EXAMPLE 1013,4-Difluoro-5-[1-(3-hydroxy-piperidin-1-yl)-methanoyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₀H₁₉F₂IN₂O₄, MS (APCI)m+1=517-   in vitro MEK assay: 75% inhibition @ 1 μM

EXAMPLE 1024,5-Difluoro-6-(4-iodo-2-methyl-phenylamino-N-(2-piperidin-1-yl-ethyl)-isophthalamicacid

-   C22H₂₄F₂IN₃O₃, MS (APCI)m+1=544-   in vitro MEK assay: 25% inhibition @ 1 μM

EXAMPLE 1033,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[2-methyl-4-(3-phenoxy-pyridin-4-yl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1042-Chloro-4-(4-{1-[2,3-difluoro-4-(4-iodo-2-methyl-phenylamino)-5-carboxy-phenyl]-methanoyl}-3-methyl-piperazin-1-yl)-benzoicacid EXAMPLE 1053,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(4-pyridin-2-yl-piperazin-1-yl)-methanoyl]-benzoicacid EXAMPLE 1065-[1-(4-Ethanesulfonyl-piperazin-1-yl)-methanoyl]-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₁H₂₂F₂IN₃O₃₅S, MS (APCI)m+1=594

EXAMPLE 1075-{1-[3-(2-Amino-ethyl)-2-oxo-imidazolidin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino-benzoicacid EXAMPLE 1085-{1-[4-(2-Amino-ethyl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1093,4-Difluoro-5-{1-[4-(2-hydroxy-2-methyl-propyl)-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1105-{1-[4-(2,4-Dimethoxy-phenyl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1115-{1-[4-(2-Carboxy-2-methyl-propyl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1123,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(1-{4-[3-(propane-1-sulfonyl)-phenyl]-piperazin-1-yl}-methanoyl)-benzoicacid EXAMPLE 1133,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(3′-methyl-2,3,5,6-tetrahydro-[1,2′]bipyrazinyl-4-yl)-methanoyl]-benzoicacid EXAMPLE 1143,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(4-{2-[(pyridin-2-ylmethyl)-amino]-ethyl}-piperazin-1-yl)-methanoyl]-benzoicacid

-   C₂₇H₂₈F₂IN₅O₃, MS (APCI)m+1=636

EXAMPLE 1155-{1-[4-(3-Dimethylamino-propyl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₄H₂₉F₂IN₄O₃, MS (APCI)m+1=587

EXAMPLE 116 3,4-Difluoro-5-{1-[4-(6-hydroxy-pyridin-2-yl)-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1175-(1-{4-[2-(2,5-Dimethyl-pyrrol-1-yl)-ethyl]-piperazin-1-yl}-methanoyl)-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1183,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(3-phenoxy-pyridin-2-yl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1193,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(5-phenoxy-pyridin-2-yl)-piperazin-1-yl]-methanoyl}-benzoicacid

-   C₃₀H₂₅F₂IN₄O₄, MS (APCI)m+1=671

EXAMPLE 1203,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(3-phenoxy-pyridin-4-yl)-[1,4]diazepan-1-yl]-methanoyl}-benzoicacid EXAMPLE 1215-{1-[4-(3-Chloro-4-hydroxymethyl-phenyl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1223,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(8-trifluoromethyl-3,4-dihydro-1H-benzo[4,5]imidazo[1,2-a]pyrazin-2-yl)-methanoyl]-benzoicacid

EXAMPLE 1235-{1-[4-(5-Chloro-1,3-dimethyl-1H-pyrazole-4-sulfonyl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₄H₂₃ClF₂IN₅O₅S, MS (APCI)m+1=694

EXAMPLE 1243,4-Difluoro-5-(1-{4-[2-(2-hydroxy-ethylsulfanyl)-phenyl]-piperazin-1-yl}-methanoyl)-2-(4-iodo-2-methyl-phenylamino)-benzoicacid p0 C₁₉H₁₇F₂IN₂O₅S, MS (APCI)m+1=551 EXAMPLE 1255-[1-(1,1-Dioxo-1⁶-thiomorpholin-4-yl)-methanoyl]-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1263,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(1-methyl-piperidin-4-yl)-piperazin-1-yl]-methanoyl}-benzoicacid

-   C₂₅H₂₉F₂IN₄O₃, MS (APCI)m+1=599

EXAMPLE 1273,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(4-oxo-1-phenyl-1,3,8-triaza-spiro[4,5]dec-8-yl)-methanoyl]-benzoicacid EXAMPLE 1283,4-Difluoro-5-{1-[4-(2-hydroxy-ethyl)-2,5-dimethyl-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₃H₂₆F₂IN₃O₄, MS (APCI)m+1=574

EXAMPLE 1293,4-Difluoro-5-{1-[4-(2-hydroxy-ethyl)-2,6-dimethyl-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE1303,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(4-isopropyl-2-methyl-piperazin-1-yl)-methanoyl]-benzoicacid EXAMPLE 1315-{1-[4-(3-Chloro-4-hydroxymethyl-phenyl)-2,6-dimethyl-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1323,4-Difluoro-5-{1-[4-(5-hydroxy-pentyl)-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1333,4-Difluoro-5-(1-{4-[2-(2-hydroxy-ethoxy)-phenyl]-piperazin-1-yl}-methanoyl)-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1343,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(4-methyl-pyridin-2-yl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1355-{1-[4-(2-sec-Butoxy-phenyl)-piperazin-1-yl]-methanoyl}3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)benzoicacid EXAMPLE 1363,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(2-isobutoxy-phenyl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1375-[1-(4-Benzothiazol-2-yl-piperazin-1-yl)-methanoyl]-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1385-{1-[4-(6-Ethoxy-pyridin-2-yl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino-benzoicacid EXAMPLE 1395-[1-(4-Benzooxazol-2-yl-piperazin-1-yl)-methanoyl]-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1403,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(3-methyl-quinoxalin-2-yl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1415-[1-(3′,6′-Dimethyl-2,3,5,6-tetrahydro-[1,2]bipyrazinyl-4-yl)-methanoyl]-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1423,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(6-methyl-pyridazin-3-yl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1433,4-Difluoro-5-(1-{4-[3-(2-hydroxy-ethoxy)-phenyl]-piperazin-1-yl}-methanoyl)-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1443,4-Difluoro-5-[1-(2-hydroxy-ethyl)-imidazolidin-2-ylidene-hydrazinocarbonyl]-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1453,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(1-{4-[3-(propane-1-sulfonyl)-phenyl]-piperazin-1-yl}-methanoyl)-benzoicacid

-   C₂₈H₂₈F₂IN₃O₅S, MS (APCI)m+1=684

EXAMPLE 1463,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(3-methanesulfonyl-phenyl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1473,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-(1-{4-[2-(propane-1-sulfonyl)-phenyl]-piperazin-1-yl}-methanoyl)-benzoicacid EXAMPLE 1485-{1-[4-(4,5-Dimethyl-thiazol-2-yl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1495-{1-[4-(5-Ethyl-[1,3,4]thiadiazol-2-yl)-piperazin-1-yl]-methanoyl}-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₃H22F₂IN₅O₃S, MS (APCI)m+1=614

EXAMPLE 1503,4-Difluoro-5-{1-[4-(1-furan-2-yl-methanoyl)-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid

-   C₂₄H₂₀F₂IN₃O₅, MS (APCI)m+1=596

EXAMPLE 1515-(1-{4-[4-(4,5-Dihydro-1H-imidazol-2-yl)-butyl]-piperazin-1-yl}-methanoyl)-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1523,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-{1-[4-(3-phosphono-propyl)-piperazin-1-yl]-methanoyl}-benzoicacid EXAMPLE 1533,4-Difluoro-5-{1-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-methanoyl}-2-(4-iodo-2-methyl-phenylamino)-benzoicacid EXAMPLE 1543,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-5-[1-(5-methyl-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-methanoyl]-benzoicacid EXAMPLE 155N-(1,1-Dioxo-tetrahydro-11⁶-thiophen-3-yl)-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-N-methyl-isophthalamicacid

-   C₂₀H₁₉F₂IN₂O₅S, MS (APCI)m+1=565

EXAMPLE 1563-({1-[5-Carboxy-2,3-difluoro-4-(4-iodo-2-methyl-phenylamino)-phenyl]-methanoyl}-amino)-tetrahydro-thiophene-3-carboxylicacid

EXAMPLE 1574,5-Difluoro-N-(1-hydroxymethyl-cyclopentyl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid EXAMPLE 1584,5-Difluoro-N-(4-hydroxy-cyclohexyl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid EXAMPLE 1594,5-Difluoro-N-((R)-2-hydroxy-cyclohexyl)-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid EXAMPLE 160N-(3-Cyclohexylamino-propyl)-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid

-   C₂₄H₂₈F₂IN₃O₃, MS (APCI)m+1=572

EXAMPLE 1614,5-Difluoro-6-(4-iodo-2-methyl-phenylamino)-N-(2-methylene-tetrahydro-thiophen-3-yl)-isophthalamicacid

-   C₁₉H₁₅F₂IN₂O₄S, MS (APCI)m+1=533

EXAMPLE 162N-(1,1-Dioxo-tetrahydro-11⁶-thiophen-3-yl)-4,5-difluoro-6-(4-iodo-2-methyl-phenylamino)-isophthalamicacid BIOLOGICAL EXAMPLES EXAMPLE 163 Cascade Assay for Inhibitors of theMAP Kinase Pathway

Incorporation of ³²P into myelin basic protein (MBP) is assayed in thepresence of a glutathione S-transferase fusion protein containing p44MAPkinase (GST-MAPK) and a glutathione S-transferase fusion proteincontaining p45MEK (GST-MEK). The assay solution contains 20 mM HEPES, pH7.4, 10 mM MgCl₂, 1 mM MnCl₂, 1 mM EGTA, 50 μM [γ-³²P]ATP, 10 μgGST-MEK, 0.5 μg GST-MAPK and 40 μg MBP in a final volume of 100 μL.Reactions are stopped after 20 minutes by addition of trichloroaceticacid and filtered through a GF/C filter mat. ³²P retained on the filtermat is determined using a 120S Betaplate. Compounds are assessed at 10μM for ability to inhibit incorporation of ³²P.

To ascertain whether compounds are inhibiting GST-MEK or GST MAPK, twoadditional protocols are employed. In the first protocol, compounds areadded to tubes containing GST-MEK, followed by addition of GST-MAPK, MBPand [γ-³²P]ATP. In the second protocol, compounds are added to tubescontaining both GST-MEK and GST-MAPK, followed by MBP and [γ-³²P]ATP.

Compounds that show activity in both protocols are scored as MAPKinhibitors, while compounds showing activity in only the first protocolare scored as MEK inhibitors.

EXAMPLE 164 In Vitro MAP Kinase Assay

Inhibitory activity can be confirmed in direct assays. For MAP kinase, 1μg GST-MAPK is incubated with 40 μg MBP in the presence or absence oftest compound for 15 minutes at 30° C. in a final volume of 50 μLcontaining 50 mM Tris (pH 7.5), 10 μM MgC1₂, 2 μM EGTA, and 10 μM[γ-³²P]ATP. The reaction is stopped by addition of Laemmli SDS samplebuffer and phosphorylated MBP resolved by electrophoresis on a 10%polyacrylamide gel. Radioactivity incorporated into MBP is determined byboth autoradiography, and scintillation counting of excised bands.

EXAMPLE 165 In Vitro MEK Assay

For evaluation of direct MEK activity, 10 μg GST-MEK, is incubated inthe presence of absence of test compound with 5 μg of a glutathioneS-transferase fusion protein containing p44MAP kinase with a lysine toalanine mutation at position 71 (GST-MAPK-KA). This mutation eliminateskinase activity of MAPK, so only kinase activity attributed to the addedMEK remains. Incubations are 15 minutes at 30° C. in a final volume of50 μL containing 50 mM Tris (pH 7.5), 10 μM MgCl₂, 2, μM EGTA, and 10 μM[γ-³²P]ATP. The reaction is stopped by addition of Laemmli SDS samplebuffer. Phosphorylated GST-MAPK-KA is resolved by electrophoresis on a10% polyacrylamide gel. Radioactivity incorporated into GST-MAPK-KA isdetermined by autoradiography, and subsequent scintillation counting ofexcised bands.

Alternatively, an artificially activated MEK containing serine toglutamate mutations at positions 218 and 222 (GST-MEK-2E) is used. Whenthese two sites are phosphorylated, MEK activity is increased.Phosphorylation of these sites can be mimicked by mutation of the serineresidues to glutamate. For this assay, 5 μg GST-MEK-2E is incubated with5 μg GST-MAPK-KA for 15 minutes at 30° C. in the same reaction buffer asdescribed above. Reactions are terminated and analyzed as above.

EXAMPLE 166 Whole Cell MAP Kinase Assay

To determine if compounds block activation of MAP kinase in whole cells,the following protocol is used. Cells are plated in multi-well platesand grown to confluence. Cells are serum-deprived overnight. Cells areexposed to the desired concentrations of compound or vehicle (DMSO) for30 minutes, followed by addition of a growth factor, for example, PDGF(100 ng/mL). After a 5-minute treatment with the growth factor, cellsare washed with PBS, and lysed in a buffer consisting of 70 mM NaCl, 10mM HEPES (pH 7.4), 50 mM glycerol phosphate, and 1% Triton X-100.Lysates are clarified by centrifugation at 13,000×g for 10 minutes. Fivemicrograms of the resulting supernatants are incubated with 10 μgmicrotubule associated protein-2 (Map2) for 15 minutes at 30° C. in afinal volume of 25 μL containing 50 mM Tris (pH 7.4), 10 mM MgCl₂, 2 mMEGTA and 30 μM [γ-³²P]ATP. Reactions are terminated by addition ofLaermmli sample buffer. Phosphorylated Map2 is resolved on 7.5%acrylamide gels and incorporated radioactivity is determined byscintillation counting of excised bands.

EXAMPLE 167 Monolayer Growth

Cells are plated into multi-well plates at 10 to 20,000 cells/mL.Forty-eight hours after seeding, test compounds are added to the cellgrowth medium and incubation is continued for 2 additional days. Cellsare then removed from the wells by incubation with trypsin andenumerated with a Coulter counter.

EXAMPLE 168

Growth in Soft-Agar

Cells are seeded into 35-mm dishes at 5 to 10,000 cells/dish usinggrowth medium containing 0.3% agar. After chilling to solidify the agar,cells are transferred to a 37° C. incubator. After 7 to 10 days' growth,visible colonies are manually enumerated with the aid of a dissectingmicroscope.

EXAMPLE 169 Collagen-Induced Arthritis in Mice

Type II collagen-induced arthritis (CIA) in mice is an experimentalmodel of arthritis that has a number of pathologic, immunologic, andgenetic features in common with rheumatoid arthritis. The disease isinduced by immunization of DBA/1 mice with 100 μg type II collagen,which is a major component of joint cartilage, delivered intradermallyin Freund's complete adjuvant. The disease susceptibility is regulatedby the class II MHC gene locus, which is analogous to the association ofrheumatoid arthritis with HLA-DR4.

A progressive and inflammatory arthritis develops in the majority ofmice immunized, characterized by paw width increases of up to 100%. Atest compound is administered to mice in a range of amounts, such as 20,60, 100, and 200 mg/kg body weight/day. The duration of the test can beseveral weeks to a few months, such as 40, 60, or 80 days. A clinicalscoring index is used to assess disease progression from erythema andedema (stage 1), joint distortion (stage 2), to joint ankylosis (stage3). The disease is variable in that it can affect one or all paws in ananimal, resulting in a total possible score of 12 for each mouse.Histopathology of an arthritic joint reveals synovitis, pannusformation, and cartilage and bone erosions. All mouse strains that aresusceptible to CIA are high antibody responders to type 11 collagen, andthere is a marked cellular response to CII.

EXAMPLE 170 SCW-induced Monoarticular Arthritis

Arthritis is induced as described by Schwab, et al., Infection andImmunity, 59:4436–4442 (1991) with minor modifications. Rats receive 6μg sonicated SCW [in 10 μl Dulbecco's PBS (DPBS)] by an intraarticularinjection into the right tibiotalar joint on day 0. On day 21, the DTHis initiated with 100 μg of SCW (250 μl) administered i.v. For oralcompound studies, compounds are suspended in vehicle (0.5%hydroxypropyl-methylcellulose/0.2% Tween 80), sonicated, andadministered twice daily (10 ml/kg volume) beginning 1 hr prior toreactivation with SCW. Compounds are administered in amounts between 10and 500 mg/kg body weight/day, such as 20, 30, 60, 100, 200, and 300mg/kg/day. Edema measurements are obtained by determining the baselinevolumes of the sensitized hindpaw before reactivation on day 21, andcomparing them with volumes at subsequent time points such as day 22,23, 24, and 25. Paw volume is determined by mercury plethysmography.

EXAMPLE 171 Mouse Ear-heart Transplant Model

Fey, T. A. et al. describe methods for transplanting split-heartneonatal cardiac grafts into the ear pinna of mice and rats (J. Pharm.and Toxic. Meth. 39:9–17 (1998)). Compounds are dissolved in solutionscontaining combinations of absolute ethanol, 0.2% hydroxypropylmethylcellulose in water, propylene glycol, cremophor, and dextrose, orother solvent or suspending vehicle. Mice are dosed orally orintraperitoneally once, twice or three times daily from the day oftransplant (day 0) through day 13 or until grafts have been rejected.Rats are dosed once, twice, or three times daily from day 0 through day13. Each animal is anesthetized and an incision is made at the base ofthe recipient ear, cutting only the dorsal epidermis and dermis. Theincision is spread open and down to the cartilage parallel to the head,and sufficiently wide to accommodate the appropriate tunneling for a rator insertion tool for a mouse. A neonatal mouse or rat pup less than 60hours old is anesthetized and cervically dislocated. The heart isremoved from the chest, rinsed with saline, bisected longitudinally witha scalpel, and rinsed with sterile saline. The donor heart fragment isplaced into the preformed tunnel with the insertion tool and air orresidual fluid is gently expressed from the tunnel with light pressure.No suturing, adhesive bonding, bandaging, or treatment with antibioticsis required.

Implants are examined at 10–20-fold magnification with a stereoscopicdissecting microscope without anesthesia. Recipients whose grafts arenot visibly beating may be anesthetized and evaluated for the presenceof electrical activity using Grass E-2 platinum subdermal pinmicroelectodes placed either in the pinna or directly into the graft anda tachograph. Implants can be examined 1–4 times a day for 10, 20, 30 ormore days. The ability of a test compound to ameliorate symptoms oftransplant rejection can be compared with a control compound such ascyclosporine, tacrolimus, or orally-administered lefluonomide.

EXAMPLE 172 Murine Ovalbumin-induced Eosinolphilia

Female C57BL/6 mice are obtained from the Jackson Laboratory (BarHarbor, Me.). All animals are given food and water ad libitum. Mice aresensitized with a single i.p. injection of OVA (grade V, Sigma ChemicalCompany, St. Louis, Mo.) adsorbed to alum, (10 μg OVA+9 mg alum in 200μl saline) or vehicle control, (9 mg alum in 200 μl saline) on day 0. Onday 14, the mice are challenged with a 12-minute inhalation of anaerosol consisting of 1.5% OVA (weight/volume) in saline produced by anebulizer (small particle generator, model SPAG-2; ICN Pharmaceuticals,Costa Mesa, Calif.). Groups of eight mice are dosed with oral vehicle(0.5% hydroxypropylmethylcellulose/0.25% TWEEN-80), or a test compoundat 10, 30, or 100 mg/kg in oral vehicle, 200 μl per mouse p.o. Dosing isperformed once per day starting on day 7 or day 13, and extendingthrough day 16.

For determination of pulmonary eosinophilia, three days after the firstOVA aerosol challenge (day 17), the mice are anesthetized with an i.p.injection of anesthetic (Ketamine/Acepromazine/Xylazine), and thetracheae is exposed and cannulated. The lungs and upper airways arelavaged twice with 0.5 ml of cold PBS. A portion (200 μl) of thebronchoalveolar lavage (BAL) fluid is enumerated using a Coulter counterModel ZB1 (Coulter Electronics, Hialeah, Fla.). The remaining BAL fluidis then centrifuged at 300×g for five minutes, and the cells areresuspended in 1 ml of HBSS (Gibco BRL) containing 0.5% fetal calf serum(HyClone) and 10 mM HEPES (Gibco BRL). The cell suspension iscentrifuged in a cytospin (Shandon Southern Instruments, Sewickley, Pa.)and stained by Diff Quick (American Scientific Products, McGraw Park,Ill.) to differentiate BAL leukocytes into neutrophil, eosinophil,monocyte or lymphocyte subsets. The number of eosinophils in the BALfluid is determined by multiplying the percentage of eosinophils by thetotal cell count.

EXAMPLE 173

Experimental compounds are added to 96 well format plates with filterbottomed wells. Kinase-inactive ERK1 (K71R mutant) in HEPES buffer isthen added to each well. After subsequent addition of MEK1 (2D mutant)diluted in a Tris buffer before being added to the plate, and thereaction is initiated by the addition of radioactive ATP, diluted in0.05% Tween 20. After 1 hour incubation at room temperature, Ice-cold20% TCA is added to each well to stop the reaction and to precipitatethe protein in solution. Filtration is done the following day followedby scintillation counting of the incorporated radioactivity using aPerkin Elmer Wallac microBeta 1450 counter. Inhibition is expressed as apercentage of the vehicle control.

From the above disclosure and examples, and from the claims below, theessential features of the invention are readily apparent. The scope ofthe invention also encompasses various modifications and adaptationswithin the knowledge of a person of ordinary skill. Examples include adisclosed compound modified by addition or removal of a protectinggroup, or an ester, pharmaceutical salt, hydrate, acid, or amide of adisclosed compound. Publications cited herein are hereby incorporated byreference in their entirety.

1. A compound of formula I:

or the pharmaceutically acceptable salts thereof; wherein R₁ ishydrogen, C₁₋₈ alkyl, C₁₋₈ alkoxy, halo, C₁₋₂ haloalkyl, or CN; R₃ andR₄ are each independently hydrogen, halo, C₁₋₂ haloalkyl, C₁₋₈ alkyl,C₁₋₈ alkoxy, nitro, CN, or (O or NH)_(k)—(CH₂)_(j)—R₉, where R₉ ishydrogen, hydroxy, CO₂H or NR₁₀R₁₁; j is 0 to 4; k is 0 or 1; R₁₀ andR₁₁ are each independently hydrogen or C₁₋₈ alkyl, or together with thenitrogen to which they are attached form a 3- to 10-member cyclic ringoptionally containing one, two, or three additional heteroatoms selectedfrom the group consisting of O, S, NH, and N—C₁₋₈ alkyl; A is hydroxy,C₁₋₆ alkoxy, or NR₆OR₇; R₆ is hydrogen, C₁₋₈ alkyl, (CO)—C₁₋₈ alkyl,phenyl, naphthyl, phenyl(C₁₋₈ alkyl), or C₃₋₁₀ cycloalkyl; R₇ ishydrogen, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₃₋₁₀ cycloalkyl orC₃₋₁₀ cycloalkyl optionally containing a heteroatom selected from thegroup consisting of O, S, and NR₉; X is NR₁₃R₁₂, or NR₁₄; R₁₂ and R₁₃are each independently hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₄₋₆ cycloalkyl, [(CH₂)_(n)Y(CH₂)_(m)]_(q)CH₃, phenyl, naphthyl, (C₁₋₆alkyl)phenyl, —[(CH₂)_(n)Y(CH₂)_(m)]_(q)phenyl, C₂₋₆ heteroaryl, (C₁₋₆alkyl)C₂₋₆ heterocyclic radical, or [(CH₂)_(n)Y(CH₂)_(m)]_(q) C₂₋₆heterocyclic radical; Y is NH or O; R₁₄ taken with N is a 5- to7-membered heterocyclic radical with between 0 and 3 additionalheteroatoms or heteroatom combinations in the ring selected from thegroup consisting of O, S, SO, SO₂, NH, and NMe;0≦n, m≦6, n+m≦8, 1≦q≦5; and wherein the above alkyl, alkenyl, alkynyl,heterocyclic radical, aryl, and cycloalkyl groups can be optionallysubstituted with between 1 and 4 substituents independently selectedfrom the group consisting of hydroxy, C₁₋₄ alkyl, fluoro, chloro, iodo,bromo, amino, and C₁₋₄ alkoxy, and NR_(a)R_(b); wherein R_(a) and R_(b)are each independently selected from the group consisting of hydrogenand C₁₋₆ alkyl.
 2. A compound of claim 1 wherein R₁ is C₁₋₈ alkyl orhalo.
 3. A compound of claim 2 wherein R₁ is methyl.
 4. A compound ofclaim 1 wherein R₃ and R₄ are each independently selected from the groupconsisting of hydrogen and halo.
 5. A compound of claim 4 wherein halois fluoro.
 6. A compound of claim 1 wherein A is NR₆OR₇.
 7. A compoundof claim 1 wherein X is NR₁₃R₁₂.
 8. A compound of claim 1 wherein X isNR₁₄.
 9. A compound of claim 1 wherein R₁₂ and R₁₃ are eachindependently selected from the group consisting of[(CH₂)_(n)Y(CH₂)_(m)]_(q)CH₃, (C₁₋₆ alkyl)phenyl,—[(CH₂)_(n)Y(CH₂)_(m)]_(q)phenyl, and (C₁₋₆ alkyl)C₂₋₆ heterocyclicradical.
 10. A compound of claim 1 wherein the heterocyclic radical is aheteroaryl selected from the group consisting of a substituted orunsubstituted radical of pyrrole, furan, pyran, thiophene, pyrazole,imidazole, triazole, tetrazole, indole, isoxazole, indazole, pyridine,pyrazine, oxazole, thiazole, oxadiazole, and oxathiadiazole.
 11. Acompound of claim 1 wherein the heterocyclic radical is a heteroalkylselected from the group consisting of a substituted or unsubstitutedradical of morpholine, piperidine, piperazine, tetrahydrofuran,tetrahydropyran, pyrrolidone, imidazoline, and tetrahydrothiophene. 12.The compound according to claim 1 where Y is O.
 13. A pharmaceuticalcomposition comprising a compound of claim 1 and a pharmaceuticallyacceptable carrier.
 14. A method of treating psoriasis in a patient inneed thereof comprising administering a therapeutically effective amountof a compound of claim
 1. 15. A method of treating osteoarthritis in apatient in need thereof comprising administering a therapeuticallyeffective amount of a compound of claim
 1. 16. A method of treatingrheumatoid arthritis in a patient in need thereof comprisingadministering a therapeutically effective amount of a compound ofclaim
 1. 17. The method of any of claims 14, 15, 16, wherein Y is O.